This data was extracted from Medline abstracts and full texts (when available) in an automated manner.
The table below describes different point mutations at a given position and provides links to other documents. The sentence(s) where the point mutations in PRIO_HUMAN at position 102 were found are listed after the table.
Yamada M, Itoh Y, Inaba A, Wada Y, Takashima M, Satoh S, Kamata T, Okeda R, Kayano T, Suematsu N, Kitamoto T, Otomo E, Matsushita M, Mizusawa H Neurology 1999 Jul 13;53(1):181-8.
Mallucci GR, Campbell TA, Dickinson A, Beck J, Holt M, Plant G, de Pauw KW, Hakin RN, Clarke CE, Howell S, Davies-Jones GA, Lawden M, Smith CM, Ince P, Ironside JW, Bridges LR, Dean A, Weeks I, Collinge J Brain 1999 Oct;122 ( Pt 10):1823-37.
De Michele G, Pocchiari M, Petraroli R, Manfredi M, Caneve G, Coppola G, Casali C, Sacca F, Piccardo P, Salvatore E, Berardelli A, Orio M, Barbieri F, Ghetti B, Filla A Can J Neurol Sci 2003 Aug;30(3):233-6.
Zanusso G, Farinazzo A, Prelli F, Fiorini M, Gelati M, Ferrari S, Righetti PG, Rizzuto N, Frangione B, Monaco S J Biol Chem 2004 Sep 10;279(37):38936-42. Epub 2004 Jul 9.
Parchi P, Chen SG, Brown P, Zou W, Capellari S, Budka H, Hainfellner J, Reyes PF, Golden GT, Hauw JJ, Gajdusek DC, Gambetti P Proc Natl Acad Sci U S A 1998 Jul 7;95(14):8322-7.
Yamada M, Tomimitsu H, Yokota T, Tomi H, Sunohara N, Mukoyama M, Itoh Y, Suematsu N, Otomo E, Okeda R, Matsushita M, Mizusawa H Neurology 1999 Jan 15;52(2):260-5.
Transgenic mice expressing the PRNP with the GSS (P102L) mutation undergo spontaneous vacuolar neurodegeneration (5)
Discussion The data in this study show that the well-studied GSS P102L mutation can cause alterations in the secondary structure and thermal properties of PrP
DISCUSSION The data in this study show that the well-studied GSS P102L mutation can cause alterations in the secondary structure and thermal properties of PrP
Reference #2 (Yamada M et al.): P102L
2 , 3 Clinically , GSS105 has been reported to be characterized by spastic paraparesis and dementia , 2 , 3 and is distinct from the classic form of GSS associated with a missense mutation at codon 102 (P102L) , which is characterized by cerebellar ataxia and dementia
Barbanti P , Fabbrini G , Salvatore M , et al. Polymorphism at codon 129 or codon 219 of PRNP and clinical heterogeneity in a previously unreported family with Gerstmann-Sträussler-Scheinker disease (PrP-P102L mutation)
Piccardo P , Ghetti B , Dickson DW , et al. Gerstmann-Sträussler-Scheinker disease (PRNP P102L) : amyloid deposits are best recognized by antibodies directed to epitopes in PrP region 90-165
Parchi P , Chen SG , Brown P , et al. Different patterns of truncated prion protein fragments correlate with distinct phenotypes in P102L Gerstmann-Sträussler-Scheinker disease
NEUROLOGY 1999 ; 53:181188 Human prion diseases are classified into three categories: infectious prion diseases , inherited prion diseases , and prion diseases of unknown etiology.1 Inherited prion diseases are found to be associated with mutations of the prion protein (PrP) gene , and they present with several clinical phenotypes , including classic and variant forms of GerstmannStra¨usslerScheinker disease (GSS) , familial CreutzfeldtJakob disease (CJD) , and familial fatal insomnia.1 A variant form of GSS associated with a missense mutation at codon 105 (P105L ; GSS105) was first described in 1993 in Japanese patients.2 , 3 Clinically , GSS105 has been reported to be characterized by spastic paraparesis and dementia , 2 , 3 and is distinct from the classic form of GSS associated with a missense mutation at codon 102 (P102L) , which is characterized by cerebellar ataxia and dementia.4 , 5 GSS105 has been referred to as the 'spastic paraparesis' type.3 Recently we experienced an autopsy case of GSS105 from a Japanese family in which we described the original patient.2 , 6 Despite the identical codon 105 mutation and codon 129 polymorphism of the PrP gene , we found remarkable differences in clinical and neuropathologic phenotypes , and detergentinsoluble PrP fragments between the family members
Reference #3 (Zanusso G et al.): P102L
In the P102L GSS variant , the ~7.5-kDa fragment has been shown to span residues 78-82 to residues 147-150 by sequence and mass spectrophotometric analyses (42 )
Singh Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L J
Reference #5 (Mallucci GR et al.): P102L
A second mutation was then described involving a proline -->leucine substitution at codon 102 (P102L) in two unrelated families with neuropathologically confirmed GSS , one British and one from the USA
The P102L mutation , absent in the normal population , was rapidly identified in numerous other unrelated GSS kindreds , including the original Austrian family described by Gerstmann (Doh-ura et al. , 1989(image) ; Brown et al. , 1991(image) ; Speer et al. , 1991(image) ; Kretzschmar et al. , 1992(image) )
Direct pathogenicity at the molecular level for the P102L mutation has been demonstrated by transgenic mice which over-express a mutant PrP with the analogous mutation to P102L in humans: P101L in mice (Hsiao et al. , 1990(image) )
It is supported by the development of neurological disease and spontaneous neurodegeneration in transgenic mice bearing the P101L mutation (equivalent to the human P102L mutation) (Hsaio et al. , 1990)
Transmissibility is a feature of all prion diseases , and transmission of the human prion diseases to laboratory primates (Brown et al. , 1994(image) ) has been successful for many cases , including the P102L mutation but not for A117V cases
It was thus proposed that different allelic forms of PRNP produce different clinical syndromes , the A117V mutation giving rise to `telencephalic GSS' and the P102L mutation giving rise to `ataxic GSS' with cerebellar pathology
As we have shown for the fourth reported family with the A117V mutation , marked variability at both the clinical and the neuropathological level is characteristic of inherited prion disease , and indeed is observed in some families with the P102L mutation , including the original Austrian family described by Gerstmann (Doh-ura et al. , 1989(image) ; Brown et al. , 1991(image) ; Speer et al. , 1991(image) ; Kretzschmar et al. , 1992(image) )
Reference #6 (Zuegg J et al.): P102L
The amino acid residues in black boxes are mutation sites known to be associated with inherited forms of PrP diseases in humans [CJD , D178N:129V (3) , V180I (3) , T183A (3) , E200K (3) , R208H (3) , V210I (3) , and M232R (3) ; GSS , P102L (3) , P105L (3) , A117V (3) , Y145Stop (3) , H187R (14) , F198S (3) , D202N (13) , Q212P (13) , and Q217R (3) ; FFI , D178N:129M (3) ; schizophrenia , N171S (12) ] , while in light gray boxes residues involved in some polymorphisms influencing these diseases are shown [M129V (3) , E219K (11) ]
Reference #7 (Inouye H et al.): P102L
Abstract X-ray diffraction was used to study the structure of assemblies formed by synthetic peptide fragments of the prion protein (PrP) that include the hydrophobic domain implicated in the Gerstmann-Sträussler-Scheinker (GSS) mutation (P102L)
The β-sheet form of Mo89-143(P101L) , carrying the equivalent of the human GSS mutation P102L , has been shown to initiate the development of GSS in transgenic mice expressing mutant PrP at low levels [Kaneko et al 2000 ]
Kirschner1 * 1 Department of Biology , Boston College , Chestnut Hill MA 02467-3811 , USA 2 Institute for Neurodegenerative Diseases and Department of Neurology , University of California , San Francisco CA 94143-0518 , USA X-ray diffraction was used to study the structure of assemblies formed by synthetic peptide fragments of the prion protein (PrP) that include the hydrophobic domain implicated in the Gerstmann-StraÈussler-Scheinker (GSS) mutation (P102L)
The b-sheet form of Mo89-143(P101L) , carrying the equivalent of the human GSS mutation P102L , has been shown to initiate the development of GSS in transgenic mice expressing mutant PrP at low levels (Kaneko et al. , 2000)
Reference #9 (Tagliavini F et al.): P102L
INTRODUCTION (image)TOP (image)ABSTRACT (image)INTRODUCTION (image)EXPERIMENTAL PROCEDURES (image)RESULTS (image)DISCUSSION (image)REFERENCES Gerstmann-Sträussler-Scheinker disease (GSS)1 is an adult-onset neurodegenerative disorder (1 , 2 ) that is inherited as an autosomal dominant trait and segregates with variant genotypes resulting from the combination of a pathogenic mutation (P102L , P105L , A117V , F198S , D202N , Q212P , and Q217R) and a common polymorphism at codon 129 (Met / Val) in the prion protein (PrP) gene (PRNP) (3-9 )
The allelic origin of the altered forms of PrP involved in the pathologic process has been investigated in other genetic prion diseases including GSS P102L , fatal familial insomnia , and Creutzfeldt-Jakob disease (CJD) associated with point or insertional mutations (19-23 )
These studies showed that only mutant PrP is detergent-insoluble and protease-resistant in GSS P102L (19 , 20 ) , fatal familial insomnia , and CJD D178N (21 ) , whereas both the mutant and wild-type proteins have these properties in CJD V210I (22 ) and CJD with five or six extra copies of the octapeptide repeat (21 )
They used a synthetic peptide homologous to mouse PrP residues 89-143 (i.e. human residues 90-144) with the P101L substitution corresponding to GSS-linked P102L mutation
Reference #10 (Chen SG et al.): P102L
We found that the main PK cleavage sites of PrP(Sc) are located at residue 97 in FFI , and residue 82 in both CJD178 and a GSS subtype linked to the P102L mutation
Reference #11 (Piccardo P et al.): P102L
3 GSS is caused by mutations P102L , P105L , A117V , G131V , F198S , D202N , Q212P , and Q217R in PRNP
Reference #12 (Spielhaupter C et al.): P102L
The location of the putative SH3 recognition motif XPXXP is indicated by an asterisk , and the two known pathologic mutations P102L and P105L in this region are indicated with arrowheads
Grb2 coprecipitates with wild type PrP-(90-231) (lane 9) as well as with the constructs bearing the GSS point mutations P102L (lane 10) and P105L (lane 11)
Such a site is present in wild type PrP at position 101-105 , but absent in two mutations leading to Gerstmann-Sträussler-Scheinker syndrome (GSS) in humans (P102L and P105L , respectively) (1 )
P102L and P105L both result in GSS with slightly different clinicopathologic patterns (50 )
Reference #13 (Kulczycki J et al.): P102L
Genetic studies disclosed in the 20th chromosome , in the PrP gene , mutation at codon 102 (P102L)
Reference #14 (Lorenz H et al.): P102L
In addition , four cellularly well defined PrP mutants (GFP-PrP P101L , GFP-PrP W144 Stop , GFP-PrP D177N , and GFP-PrP E199K as the mouse homologues for human PrP P102L , Y145 Stop , D178N , E200K) were used as controls to evaluate the GFP-PrP model system and to extend the existing knowledge about these mutants (7 , 17 , 19 , 20 )
Biochemical as well as cellular data have been described for four of the seven disease-related PrP mutants , i.e. the three full-length PrP mutants PrP P102L , PrP D178N , PrP E200K (7 , 19 ) and the truncated molecule PrP Y145 Stop (20 ) but not for PrP Q160 Stop , PrP T188K , and PrP T188R (human PrPs)
GFP-PrP chimera with mouse PrPa Human PrP Phenotype P101L P102L GSS W144Stopb Y145Stop GSS Q159Stop Q160Stop EODc D177N D178N FFI T187K T188K CJD T187R T188R CJDd E199K E200K CJD a Mouse PrP contains methionine at position 128 , which corresponds to codon 129 in human PrP , a polymorphic site encoding either methionine or valine
Reference #15 (Mastrianni JA et al.): P102L
PRNP mutations appear to be associated with distinct clinicopathologic phenotypes such as the ataxic presentation of Gerstmann-Sträussler-Scheinker disease associated with the P102L mutation 1 -3 and the insomnia and dysautonomia of the D178N , M129 haplotype , which determines fatal familial insomnia (FFI)
Reference #16 (Wegner C et al.): P102L
Therefore , we did not find any indication of a defect in the post-translational transport of the mutant PrPs to the cell surface , whereas a few pathogenic human mutations , such as P102L , D178N and F197 , as well as an insertional mutation with nine additional octapeptides , were shown to have an effect on the delivery of the molecules to the cell surface (Ivanova et al. , 2001(image) )
In the latter study , the authors found that , under such conditions , the introduction of the human-pathogenic mutations P102L , D178N , T183A and E200K and an insertion of six additional octapeptides in the ORF of Prnp at the respective locations (P101L , D177N , T183A and E199K) rendered the mutated proteins resistant to PK at the above concentration
Reference #17 (Mishra RS et al.): P102L
Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L* Received for publication , January 8 , 2002 , and in revised form , April 11 , 2002 Published , JBC Papers in Press , April 19 , 2002 , DOI 10.1074 / jbc.M200213200 Ravi Shankar Mishra , Yaping Gu , Sharmila Bose , Susamma Verghese , Sudheera Kalepu , and Neena Singh From the Institute of Pathology , Case Western Reserve University , Cleveland , Ohio 44106 A familial prion disorder with a proline to leucine substitution at residue 102 of the prion protein (PrP102L) is typically associated with protease-resistant PrP fragments (PrPSc) in the brain parenchyma that are infectious to recipient animals
Alternate mice expressing a single copy of PrP102L (mouse PrP101L) do not develop spontaneous disease , but show dramatic susceptibility to PrPSc isolates from different species
To understand these discrepant results , we studied the biogenesis of human PrP102L in a cell model
Here , we report that cells expressing PrP102L show decreased expression of the normal 18-kDa fragment on the plasma membrane
Thus , altered susceptibility of PrP101L mice to exogenous PrPSc may be mediated by the 20-kDa CtmPrP fragment , rather than PrP102L per se
Thus , it appears that PrP102L alters certain cellular characteristics or functions of neuronal cells that influence the replication or toxicity of exogenously introduced PrPSc
We investigated the biogenesis of PrP102L in transfected human neuroblastoma cells in an attempt to model the events that might occur in vivo , and uncover the biochemical pathways of neurotoxicity in GSS102L
In this report , we show that the metabolism of PrP102L is altered , resulting in accumulation of a 20-kDa fragment of PrP on the cell surface , with a concomitant decrease in the expression of 18-kDa fragment , a product of normal recycling of PrP from the plasma membrane (15)
Our results suggest that the change in ratio of 18:20-kDa fragments on PrP102L cells may increase their vulnerability to PrPSc toxicity by unconventional pathways , thus accounting for the complex biological effects of PrP102L in vivo
1 The abbreviations used are: GSS , Gerstmann-Straussler-Scheinker disease ; PrPM , mutant PrP ; PrPC , normal cell-associated PrP ; PrPSc , conformationally transformed scrapie form of PrP ; CtmPrP , transmembrane PrP with the C terminus in the endoplasmic reticulum lumen ; NtmPrP , transmembrane PrP with the N terminus in the ER lumen ; PrP102L , PrP with a Pro to Leu mutation at codon 102 of the prion protein ; PK , proteinase K ; PNGase-F , N-glycosidase F ; GPI , glycosyl phosphatidylinositol ; PI-PLC , phosphatidylinositol-specific phospholipase C ; ER , endoplasmic reticulum ; FITC , fluorescein isothiocyanate ; PBS , phosphate-buffered saline ; TBS , Tris-buffered saline ; DMEM , Dulbecco's modified Eagle's medium ; PMSF , phenylmethylsulfonyl fluoride ; TRITC , tetramethyl rhodamine isothiocyanate
Transfected M17 cells expressing wild type (PrPC) or mutant (PrP102L) prion protein were generated as described in a previous report (17 , 18)
Equal amount of total protein was used from cells expressing either PrPC or PrP102L
Metabolic Labeling and Immunoprecipitation--In a typical experiment , 9 106 cells expressing PrPC or PrP102L were used
Labeling at 15 °C--Two 10-cm dishes containing 9 106 PrPC- or PrP102L-expressing cells were washed with methionine-cysteine free DMEM , and preincubated in the same medium containing 5% dialyzed serum for 1 h at 37 °C
Cell Homogenization and Treatment with Proteinase K--PrPC- and PrP102L-expressing cells were washed and homogenized on ice in a buffer containing 10 mM HEPES , pH 7.9 , 1.5 mM MgCl2 , 10 mM KCl , and 0.5 mM dithiothreitol with 20 strokes of a Kontes all-glass Dounce homogenizer
Assay of PrP Endocytosis--Cells expressing PrPC or PrP102L were cultured on poly-D-lysine-coated glass coverslips overnight
One set of PrPC and PrP102L cells were incubated with anti-PrP antibody 3F4 (1:25) in complete medium for 30 min on ice , and two other sets of cells were incubated with the same antibody at the same dilution for 10 or 60 min each at 37 °C in a humidified CO2 incubator
RESULTS Surface Expression of 18- and 20-kDa C-terminal Fragments of PrP Is Altered in PrP102L Cells--The steady state expression of PrPC and PrP102L in transfected neuroblastoma cells was evaluated by immunoblotting cell-associated PrP with antibodies specific to PrP residues 109 and 112 (3F4) , 145180 (8H4) , or 220 230 (2301)
Lysates of PrP102L showed similar glycoforms of PrP and the 20-kDa fragment , but in contrast to PrPC , all the PrP102L glycoforms and the 20-kDa fragment migrated 1 kDa faster on SDS-PAGE (Fig. 1 , lanes 1 and 2 versus lanes -->and 4)
More importantly , the 20-kDa fragment is 4-fold more in PrP102L lysates as compared with PrPC (lane 2 versus lane 4)
Similar processing of PrP102L lysates revealed the three glycoforms of PrP , but surprisingly , the 18-kDa fragment and its glycosylated form of 22 kDa were barely detectable (Fig. 1 , lane 7)
However , as compared with PrPC , the 18-kDa fragment was 5-fold less , and the 20-kDa fragment 4-fold more in PrP102L lysates
Unlike the 20-kDa fragment , migration of 18-kDa fragment of PrPC and PrP102L was similar (Fig. 1 , lanes 6 and 8)
The increased representation of 20-kDa fragment in PrP102L lysates could be the result of increased production resulting from abnormal metabolism of PrP102L or , conversely , decreased turnover of the 20-kDa fragment caused by misfolding or aggregation because it probably includes the PrP102L mutation
The decreased representation of 18-kDa fragment , on the other hand , could be caused by reduced expression of full-length PrP102L on the cell surface because of sequestration in an intracellular compartment or , conversely , normal surface expression but decreased endocytosis and / or recycling to the plasma membrane
Over-representation of 20-kDa Fragment in PrP102L Cells Is Not the Result of Increased Production--To evaluate whether accumulation of the 20-kDa fragment is the result of increased production as a result of aberrant metabolism of PrP102L , PrPCand PrP102L-expressing cells were labeled with Tran35S-label for 2 h and subjected to immunoprecipitation with 3F4 or 8H4 antibodies
The three glycoforms of PrP and the 20-kDa fragment were detected with 3F4 in both PrPC and PrP102L lysates , and as observed in Fig. 1 , PrP102L glycoforms and the 20-kDa fragment migrated 1 kDa faster than PrPC forms (Fig. 2A , lanes 1 and 2)
The 20-kDa fragment was prominent in both samples , but in contrast to the results obtained in Fig. 1 at steady state , the 20-kDa fragment was not over-represented in PrP102L lysates (Fig. 2A , lanes 1 and 2)
A small amount of 18-kDa fragment could be detected in 8H4 immunoprecipitates , but there was no significant difference in the amount detected in PrPC and PrP102L lysates (Fig. 2A , lanes 3 and 4)
Because the difference in the amount of 18- and 20-kDa fragments in PrPC and PrP102L lysates was detected at steady state but not after a 2-h pulse , it probably arose as a result of a cumulative process that becomes apparent over time , and not because of an acute abnormality in the metabolism of PrP102L
To confirm the above results , the synthesis and turnover of PrPC and PrP102L were compared in a pulse-chase paradigm
PrP102L samples showed similar kinetics of synthesis and transport of full-length and 20-kDa forms to the plasma membrane (Fig. 2B , lanes 4 6 and 10 12) , except for a significant decrease in full-length PrP102L forms after 4 h of chase both in the lysate and PI-PLC-cleaved samples (Fig. 2B , lanes 6 and 12)
In addition , a 14-kDa form was detected in the PrP102L lysates soon after the pulse , which decreased gradually with chase (Fig. 2B , lanes 4 6)
Although the 14-kDa form of PrP102L was not detected in the PI-PLC-cleaved samples , subsequent experiments with long term labeling showed that it was indeed secreted into the culture medium (see Fig. 4B)
Surface expression of 18- and 20-kDa C-terminal fragments of PrP is altered in PrP102L cells
Immunoblotting of PrPC and PrP102L lysates with 3F4 shows three glycoforms of PrPC comprising the unglycosylated (27 kDa) , intermediate (29 30 kDa) , and highly glycosylated forms (33 42 kDa) (lane 1)
PrP102L lysates show similar glycoforms of PrP and the 20-kDa fragment , but in contrast to PrPC , the 20-kDa fragment is 4 times more in PrP102L lysates (lane 2 versus lane 4)
In addition , all the PrP102L glycoforms and the 20-kDa fragment migrate 1 kDa faster than PrPC on SDS-PAGE (lanes 1 and 2 versus lanes -->and 4)
PrP102L lysate , on the other hand , reveals the three PrP glycoforms , but the 18-kDa fragment and its 22-kDa glycosylated form are barely detectable (lane 7)
Deglycosylation shows the 27-kDa full-length form as above , but in contrast to PrPC , the 18-kDa fragment is 5-fold less , and the 20-kDa is 4-fold more in PrP102L lysates (lane 8)
A stronger 20-kDa band is observed in 3F4 blots because of higher affinity of 3F4 as compared with 8H4 antibody Accumulation of Truncated CtmPrP in GSS P102L arises from an alternate form of PrP , probably the transmembrane PrP (CtmPrP) , and the accumulation of this fragment observed in PrP102L lysates in Fig. 1 is not the result of increased production or sequestration in an intracellular compartment , but perhaps of reduced degradation
To check this assumption , PrPC and PrP102L cells labeled overnight with the anchor component [3H]ethanolamine were lysed , immunoprecipitated with 3F4 , and treated with PNGase-F to remove all glycans
The samples were fractionated on a long SDS-PAGE gel to accentuate the difference in migration of the 20-kDa fragment from PrPC and PrP102L lysates
As expected , the 27-kDa full-length form and the 20kDa fragment of PrP102L migrated 1 kDa faster on SDSPAGE (Fig. 2C , lane 1 versus lane 2)
More importantly , the 20-kDa fragment from both PrPC and PrP102L lysates was labeled with [3H]ethanolamine , confirming that it is linked with the GPI anchor (Fig. 2C , lanes 1 and 2)
As observed in Fig. 1 above , the 20-kDa was 4-fold more in PrP102L lysates as compared with PrPC
The 20-kDa Fragment Is Generated from Transmembrane PrP in the Endoplasmic Reticulum--To evaluate whether the 20-kDa fragment arises from CtmPrP , PrPC and PrP102L cells were treated with the proteasomal inhibitor MG132 for 2 h , and subjected to immunoblotting with 3F4
To further confirm the origin of 20-kDa fragment from CtmPrP , microsomes prepared from PrPC and PrP102L cells were treated with 20 g / ml PK on ice for 30 min and subjected to immunoblotting with 3F4 or anti-calnexin antibodies
As shown in Fig. 3B , there was a small but significant increase in the 20-kDa fragment after protease digestion of PrPC and PrP102L microsomes , whereas the full-length PrP glycoforms were virtually unchanged (Fig. 3B , lane 1 versus lane 2 and lane -->versus lane 4)
Similar analysis of cell homogenates prepared from cells cultured with the proteasomal inhibitor MG132 for 2 h showed increased expression of the 26- and 20-kDa fragments in both PrPC and PrP102L preparations (Fig. 3B , lanes 5 8)
PrP102L samples , in addition , showed a doublet in the region of 14 kDa when proteasomal function was inhibited (Fig. 3B , lane 7)
PK treatment of microsomes prepared from these cells showed an increase in the 20-kDa fragment in both PrPC and PrP102L samples , whereas the 26-kDa fragment and full-length PrP forms were unaffected (Fig. 3B , lane 5 versus lane 6 and lane 7 versus lane 8)
To evaluate whether up-regulation of CtmPrP synthesis in PrP102L cells accounts for the over-representation of 20-kDa fragment , PrPC and PrP102L cells were radiolabeled in the FIG
Over-representation of 20-kDa fragment in PrP102L cells is not the result of increased production
A , PrPC and PrP102Lexpressing cells were labeled with Trans35S-label for 2 h and subjected to immunoprecipitation with 3F4 or 8H4
The three glycoforms of PrP and the 20-kDa fragment are detected with 3F4 in both PrPC and PrP102L lysates (lanes 1 and 2) , and , as observed in Fig. 1 , PrP102L glycoforms and the 20-kDa fragment migrate 1 kDa faster than PrPC forms
The 20-kDa fragment is prominent in both samples , but is not over-represented in PrP102L lysates (lanes 1 and 2)
B , PrPC and PrP102L cells were labeled with Tran35S-label for 30 min and chased for 0 , 2 , and 4 h
PrP102L samples show similar kinetics of synthesis and transport of full-length and 20-kDa forms to the plasma membrane (lanes 4 6 and 10 12)
Note the decrease in full-length PrP102L after 4 h of chase in the lysate and PI-PLC-cleaved samples (lanes 6 and 12)
In addition a 14-kDa form is detected in the PrP102L lysate soon after pulse , which decreases gradually with chase (lanes 4 6)
C , PrPC and PrP102L cells were labeled overnight with the anchor component [3H]ethanolamine , subjected to immunoprecipitation with 3F4 , and deglycosylated
As expected , the 27-kDa full-length form and the 20-kDa fragment of PrP102L migrate 1 kDa faster on SDS-PAGE (lane 1 versus lane 2)
The 20-kDa fragment from both PrPC and PrP102L lysates is labeled with [3H]ethanolamine (lanes 1 and 2)
The 20-kDa fragment was detected even at 15 °C in both PrPC and PrP102L lysates , confirming that it is generated in the ER (Fig. 3C , lanes 2 and 4)
Surprisingly , an additional fragment of 14 kDa was detected at 15 °C in PrP102L lysates (Fig. 3C , lane 4)
Quantitative estimation of the above results in terms of percentage increase of 20 kDa in comparison to full-length PrP forms shows increased accumulation of the 20-kDa fragment at 15 °C , and in the presence of MG132 in both PrPC and PrP102L lysates
The percentage of 20 kDa in PrP102L lysates was less than PrPC under all four conditions examined , i.e. at 37 °C or 15 °C , and in the absence or presence of MG132
The 14-kDa fragment appears to be highly unstable , because it was detected only at 15 °C or in the presence of MG132 and was significantly increased in PrP102L cells
Together , the above results show that: 1) the 20-kDa fragment is generated in the ER , most likely from proteolytic cleavage of CtmPrP at the ER membrane , 2) inhibition of CtmPrP degradation by proteasomal inhibition results in an increase in the generation of 20-kDa fragment , 3) an additional N-terminal fragment of 14 kDa is detected in PrP102L cells , the identity of which is presently unclear , and 4) increased representation of 20-kDa fragment in PrP102L is not because of increased synthesis of CtmPrP , but rather because of decreased turnover of this fragment , probably at the cell surface
In the following experiments , cell surface expression and turnover of the 20kDa fragment were investigated to understand the cellular processes that lead to its accumulation in PrP102L cells
The 20-kDa Fragment Accumulates on the Surface of PrP102L Cells--As noted in Fig. 1 , not only is the 20-kDa over-represented in PrP102L at steady state , but the 18-kDa fragment of PrP is significantly decreased
To study the metabolism of 18and 20-kDa fragments under conditions resembling steady state , PrPC- and PrP102L-expressing cells were labeled with Tran35S-label overnight in a 3:1 ratio of methionine-cysteinefree and normal DMEM containing 5% dialyzed serum
PrP102L lysates showed under-representation of the unglycosylated form as compared with PrPC , and all the PrP102L forms migrated 1 kDa faster than PrPC (Fig. 4A , lane 2)
As observed in the PrPC samples , the 20-kDa fragment was mostly detected in the PI-PLC-cleaved material , and was over-represented in PrP102L as compared with PrPC (Fig. 4A , lanes -->and 4)
A , PrPC and PrP102L cells were cultured in the presence of the proteasomal inhibitor MG132 for 2 h and subjected to immunoblotting with 3F4
B , microsomes prepared from PrPC and PrP102L cells were treated with 20 g / ml PK on ice for 30 min and subjected to immunoblotting with 3F4 or anti-calnexin antibody
A significant increase in the 20-kDa fragment is observed after protease digestion of PrPC and PrP102L microsomes , whereas the full-length PrP glycoforms are virtually unchanged (lane 1 versus lane 2 and lane 3 versus lane 4)
PrPC and PrP102L cells treated with MG132 for 2 h show accumulation of 26-kDa (arrow) and 20-kDa forms (lanes 5 8) , and a 14-kDa species in PrP102L lysates (lane 7)
C , PrPC or PrP102L cells were radiolabeled in the absence or presence of the proteasomal inhibitor MG132 for 2 h at 37 °C , or at 15 °C to block transport of proteins from the ER
The 20-kDa fragment is detected even at 15 °C in both PrPC and PrP102L lysates , confirming that it is generated in the ER (lanes 2 and 4)
An additional fragment of 14 kDa is detected at 15 °C in PrP102L lysates (lane 4)
D , quantitative estimation of the above results in terms of percentage increase of 20 kDa in comparison to full-length PrP forms shows increased accumulation of 20-kDa fragment at 15 °C , and in the presence of MG132 in both PrPC and PrP102L lysates
Accumulation of Truncated CtmPrP in GSS P102L PrP102L samples became more prominent following deglycosylation of PI-PLC-cleaved samples , indicating that a significant proportion of this fragment on the cell surface is glycosylated , and migrates at 20 kDa when glycans are removed (Fig. 4A , lanes 5 and 6)
In addition , a 14-kDa fragment was detected in PI-PLC-cleaved proteins obtained from PrP102L cells (Fig. 4A , lane 4)
Immunoprecipitation with 8H4 showed similar PrPC and PrP102L glycoforms as observed with 3F4
As noted in Fig. 1 , the 18-kDa fragment was under-represented and the 20-kDa fragment was over-represented in PrP102L lysates (Fig. 4A , lanes 8 and 10)
Deglycosylation of PI-PLC cleaved samples showed 5-fold less 18-kDa and 4-fold more 20-kDa fragment in PrP102L compared with PrPC (Fig. 4A , lanes 11 and 12)
Immunoprecipitation of medium collected from PrPC and PrP102L cells with 3F4 showed the presence of 14-kDa fragment in PrP102L (Fig. 4B , lane 2) , indicating that this fragment is secreted into the medium
A small amount of the full-length PrPC and PrP102L forms was also shed into the medium (Fig. 4B , lanes 1 and 2)
Thus , at steady state (Fig. 1) or after prolonged radiolabeling , two significant differences were observed in PrP102L: 1) the C-terminal 18-kDa fragment is significantly decreased , and 2) the C-terminal 20-kDa fragment is increased in PrP102L cells
From the pulse-chase experiments , it is clear that PrP102L was transported to the plasma membrane normally
The marked decrease in 18-kDa may therefore arise because of inefficient endocytosis or recycling of PrP102L at the plasma membrane
In addition , it is clear from the above data that the over-representation of 20 kDa in PrP102L is not the result of up-regulation of CtmPrP , or of an increase in the proteolytic processing of CtmPrP
Endocytosed PrP102L Is Targeted to Lysosomes Instead of Recycling Back to the Plasma Membrane--The kinetics of PrPC and PrP102L endocytosis and recycling were examined by immunofluorescence analysis
PrPC and PrP102L cells were incubated with anti-PrP antibody 3F4 in complete culture medium for 30 min on ice , or for 10 and 60 min each at 37 °C in a CO2 incubator
Following 30 min of incubation with 3F4 on ice , almost all of the PrPC and PrP102L were detected on the cell surface (Fig. 5A , red , panels 1 and 2)
In contrast , the intracellular vesicles in PrP102L were concentrated in a perinuclear location rather than close to the plasma membrane (Fig. 5A , panel 4) , a difference that became more apparent after 60 min of incubation with 3F4 (Fig. 5A , panel 5 versus panel 6)
Fig. 5B shows confocal images of the same experiment at a higher magnification to emphasize the difference between intracellular localization of endocytic vesicles loaded with PrPC (Fig. 5B , panels 1 , 3 , and 5) or PrP102L (Fig. 5B , panels 2 , 4 , and 6)
In addition , there are significant differences in the fluorescence intensity of PrPC and PrP102L at the cell surface
After 10 min of incubation at 37 °C , the surface expression of PrPC and PrP102L is similar (Fig. 5 , A and B , panels -->and 4)
However , after 60 min , the surface expression of PrPC and PrP102L was significantly different
In PrP102L cells , only the red stain was prominent , suggesting that a fair amount of PrP102L did not get internalized even after 1 h of incubation at 37 °C , and that internalized PrP102L-antibody complexes that should have stained green did not resurface back to the plasma membrane (Fig. 5 , A and B , panels 5 and 6)
To check whether the reduced surface expression of PrP102L is caused by targeting of internalized molecules to the lysosomes , PrPC and PrP102L cells incubated with 3F4 for 60 min as above were processed for co-immunostaining with cathepsin-D , a marker for late endosomes and lysosomes , or LysoTracker , a marker for lysosomes
As opposed to PrPC , a significant amount of co-staining was observed in PrP102L samples (data not shown)
Together , the above data indicate that , as opposed to PrPC , PrP102L is probably degraded by the lysosomes following endocytosis and not recycled back to the plasma membrane
The 20-kDa fragment accumulates on the surface of PrP102L cells
A , PrPC and PrP102L cells were labeled with Tran35Slabel overnight in a 4:1 ratio of methionine-cysteine-free and normal DMEM containing 5% dialyzed serum
PrP102L lysates show under-representation of the unglycosylated form as compared with PrPC , and all the PrP102L forms migrate 1 kDa faster than PrPC (lanes 1 and 2)
The 20-kDa fragment is more prominent in the PI-PLCcleaved sample and is over-represented in PrP102L as compared with PrPC (lanes -->and 4)
Deglycosylation with PNGase-F accentuates the difference in the amount of 20-kDa between PrPC and PrP102L samples (lanes 5 and 6)
An additional band of 14 kDa is detected in the PI-PLC-cleaved sample of PrP102L (lane 4)
Immunoprecipitation with 8H4 shows similar PrPC and PrP102L glycoforms as observed with 3F4 (lanes 7 and 8)
The 18-kDa fragment is under-represented , and the 20-kDa fragment is over-represented , in PrP102L lysates (lanes 8 and 10)
Deglycosylation of PI-PLC cleaved samples accentuates the difference in 18and 20-kDa fragments in PrPC and PrP102L (lanes 11 and 12)
B , medium collected from overnight labeling of PrPC and PrP102L cells was immunoprecipitated with 3F4 and analyzed
A small amount of the full-length PrPC and PrP102L forms (lanes 1 and 2) and a 14-kDa fragment are detected in PrP102L samples (lane 2)
In this report , we provide a detailed analysis of the metabolic consequences of PrP102L mutation in transfected neuroblastoma cells
We show that the processing and turnover of PrP102L is altered , resulting in decreased expression of the normal 18-kDa fragment , and increased accumulation of the 20-kDa CtmPrP fragment on the surface of these cells
This change in phenotype may render PrP102L cells more susceptible to exogenous PrPSc infection and toxicity by two inter-related mechanisms
In our cell model of PrP102L , the surface representation of 20-kDa CtmPrP fragment is 4-fold more , and the 18-kDa 5-fold less , as compared with PrPC cells
We believe that this difference arises from aberrant recycling of PrP102L from the cell surface
Because the 20-kDa fragment , but not the 18kDa fragment , includes most of this region , a change in the ratio of 20- to 18-kDa fragment on the surface of PrP102Lexpressing cells could have profound implications for PrPSc replication and neurotoxicity in vivo
In our cell model , PrP102L is neither aggregated nor PKresistant
Endocytosed PrP102L is targeted to lysosomes instead of recycling back to the plasma membrane
PrPC and PrP102L cells were incubated with anti-PrP antibody 3F4 in complete culture medium for 30 min on ice or for 10 and 60 min at 37 °C under normal culture conditions
A , following 30 min of incubation with 3F4 on ice , almost all of the PrPC and PrP102L are detected on the cell surface (panels 1 and 2)
In contrast , intracellular vesicles in PrP102L are concentrated in a perinuclear location (panel 4)
B , confocal images of the above experiment at a higher magnification to emphasize the difference in intracellular localization of PrPC (panels 1 , 3 , and 5) and PrP102L (panels 2 , 4 , and 6)
After 60 min of incubation with 3F4 the surface expression of PrPC is significantly more than PrP102L (panels 5 and 6)
Accumulation of Truncated CtmPrP in GSS P102L plify the neurotoxic signal initiated by exogenous PrPSc without itself undergoing a transformational change , causing neurotoxicity by a CtmPrP-mediated phenomenon
Although PK treatment of microsomes prepared from PrPC and PrP102L cells resulted in an increase in the amount of 20-kDa fragment , this result only confirms the origin of 20-kDa fragment from CtmPrP
The amount of 20-kDa fragment generated in the two cell lines is not altered significantly , and thus the increased representation of 20-kDa fragment in PrP102L cells cannot be attributed to an up-regulation in the synthesis , or increased proteolytic cleavage of CtmPrP
Instead , we believe that the accumulation of 20-kDa fragment on the surface of PrP102L cells is caused by decreased turnover
Our results on the kinetics of endocytosis and recycling of PrPC and PrP102L show clearly that , whereas PrPC remains in peripheral endosomes close to the plasma membrane and is recycled back to the cell surface , most of PrP102L accumulates in perinuclear structures , probably lysosomes , and is degraded
These results explain the decreased expression of 18-kDa fragment on the surface of PrP102L cells , because 18-kDa fragment must recycle back to the plasma membrane after cleavage in an endocytic compartment
The mechanism leading to over-representation of 20-kDa fragment on the surface of PrP102L cells is not entirely clear from our data
Because the 20-kDa fragment in PrP102L cells migrates faster on SDS-PAGE than the corresponding fragment from PrPC , it probably includes the PrP102L mutation
The faster migration in the 20-kDa and not the 18-kDa fragment of PrP102L is probably a result of the amino acid change to leucine , allowing better binding to SDS than proline , which is a helix breaker
It is possible that the 20-kDa fragment with the PrP102L mutation is endocytosed less efficiently because of the mutation , which may cause a change in its conformation
A similar change in secondary structure may account for the lysosomal delivery of PrP102L after endocytosis rather than recycling back to the plasma membrane
Such an alteration in the targeting and turnover of full-length PrP102L and its 20kDa fragment would reverse the ratio of 20- to 18-kDa fragment on the surface of these cells , thus altering their phenotype in a significant way
However , both full-length PrP102L and its 20-kDa fragment can be released from the cell surface by PI-PLC , excluding the possibility that these are grossly misfolded or aggregated on the cell surface
In conclusion , this report highlights important differences in the metabolism of PrP102L as compared with PrPC , implicating the 20-kDa metabolic product of CtmPrP in the pathogenesis of GSS102L
Reference #18 (Mastrangelo P et al.): P102L
In the case of GSS it was not possible to undertake meaningful analyses for the causative P102L , P105L and A117V mutations as these are located within an area of PrP that has no cognate in Dpl [16 ]
Reference #19 (Cohen E et al.): P102L
Because there is an abundance of prolines in PrP , and since two proline substitutions (P102L and P105L) are linked with familial GSS (Hsiao et al. , 1989 ; Yamazaki et al. , 1999 ) , we wondered whether cis-trans isomerization of peptidylprolyl bonds might participate either in the folding of PrP C or in its disposal
(B) Model for the generation of misfolded PrP by wtPrP and by the P102L and P105L mutants
Upper panel: in all cases the majority of both wild-type and mutant (P102L and P105L) PrP is formed with predominantly trans X-Pro (or trans X-Leu) peptide bonds (which are energetically favored)
The substitutions P102L and P105L are linked , separately , to familial GSS (Hsiao et al. , 1989 ; Yamazaki et al. , 1999 ) , but the etiology is poorly understood
The biochemical properties of MoP101L-PrP (the mouse homolog of HuP102L-PrP) have been characterized by Harris and colleagues , who have shown that this mutant is slightly protease resistant (Lehmann and Harris , 1995 )
Although this could apply to any of the X-Pro bonds in mature wild-type huPrP , it is tempting to focus on P102 and P105 , the two proline residues that are linked to familial GSS [P102L (Hsiao et al. , 1989 ) ; P105L (Yamazaki et al. , 1999 )]
It is thus plausible that P102L- and P105L-PrP are sometimes (even if rarely) synthesized with cis X-Leu bonds at positions 102 or 105
(i) It forecasts that while a small number of P102L or P105L PrP molecules may acquire prion-like properties , the vast majority of these mutant molecules will form in the native conformation
Reference #20 (Sasaki K et al.): P102L
Gerstmann-Straussler-Scheinker disease (GSS) is a hereditary transmissible spongiform encephalopathy associated with prion protein gene mutation P102L
We had a chance to examine an autopsy case with PRNP P102L mutation
Reference #21 (Barron RM et al.): P102L
This line of mice was generated to model the mutation thought to be responsible for P102L GSS , a familial TSE disease in humans
Reference #22 (De Michele G et al.): P102L
Molecular analysis showed P102L mutation in PRNP gene
Reference #23 (Fraser E et al.): P102L
The relationships between the degree of cortical prion protein (PrP) deposition , tissue vacuolation and astrocytosis were studied in the frontal cortex of 27 cases of human spongiform encephalopathy , encompassing 13 cases of sporadic Creutzfeldt-Jakob disease (sCJD) , four cases of familial CJD (fCJD) (one owing to E200K mutation , one owing to 144 bp insertion , one owing to P102L mutation and one owing to A117V mutation) , five cases of iatrogenic CJD (iCJD) owing to growth hormone therapy and five cases of variant CJD (vCJD)
Reference #24 (Bianca M et al.): P102L
Case report Gerstmann– ; Sträussler– ; Scheinker disease with P102L– ; V129 mutation: a case with psychiatric manifestations at onset Marco Biancaa , Sebastiano BiancaCorresponding Author Contact Information , E-mail The Corresponding Author , b , Ignazio Vecchioa , Rocco Raffaelea , Carmela Ingegnosib and Francesco Nicolettia a Neurosciences Department , University of Catania , Via S
We describe a patient with GSS and P102L– ; V129 mutation in which the onset with prominent psychiatric features characterized by apathy and depression and not with cerebellar sign and the clinical course with seizures , nor observed in P102L– ; V129 cases , allow us to confirm observations that the GSS caused by the 102 mutation is influenced by the codon 129 polymorphism with a specific genotype– ; phenotype influence , but probably other additional factors might be considered as background for phenotypic variability
The most common mutation causing GSS disease is P102L– ; V129 , which results in the substitution of proline to leucine in coupling with methionine at residue 129 [5 ]
Mutation at P102L with valine at 129 has also been reported [8 ]
We describe a patient with GSS and P102L– ; V129 mutation focused on possible genotype– ; phenotype correlations
A P102L– ; V129 mutation of PRNP gene was detected
In fact the clinical course observed in patients with P102L– ; V129 is significantly different from that observed in P102L– ; V129 patients that showed an age of onset in the fourth to seventh decade of life , and the duration of disease ranges from 1 to 10 years
In fact the presentation was with prominent psychiatric features characterized by apathy and depression and not with cerebellar sign that appeared only later , also seizures were not observed in P102L– ; V129 cases , but were reported in P102L– ; V129 [8 ]
Parchi et al. [9 ] demonstrated that the heterogeneity of the clinico-pathologic phenotype associated with GSS P102L variant is related to the presence and relative abundance of two protease-resistant PrP fragments (PrP-res) that have different size and degree of glycosilation
Reference #25 (Tremblay P et al.): P102L
One mutation causing GSS is P102L , denoted P101L in mouse PrP (MoPrP)
In earlier studies , transgenic (Tg) mice , denoted Tg2866 , expressing high levels of PrP(P101L) were used to model Gerstmann-Stra¨ussler-Scheinker (GSS) disease caused by the P102L point mutation
In a limited study , GSS(P102L) was transmitted to Tg mice expressing a chimeric mouse-human (MHu2 M) PrP transgene carrying the P102L mutation but not * Corresponding author
In another study , GSS(P102L) human prions were transmitted to Tg mice expressing MoPrP(P101L) in which the transgene was incorporated through gene replacement (31)
The results reported here were derived from studies using Tg(MoPrP , P101L)196 / Prnp0 / 0 , Tg(MoPrP , P101L)2866 / Prnp0 / 0 , Tg(MoPrP , P101L)2247 / Prnp0 / 0 , Tg(MoPrP)4053 / Prnp / , Tg(MHu2M)5378 / Prnp0 / 0 , and Tg(MHu2MPrP , P102L)69 / Prnp0 / 0 mice ; these Tg lines are described in detail elsewhere (27 , 47 , 48)
The serum from rabbit 5449 displayed strong reactivity against MoPrP(P101L) , MHu2M(P102L) , and HuPrP(P102L) but not against wt controls (Fig. 1)
Brain homogenates (20 g of total protein per lane) from mice expressing wt MoPrP-A (FVB) , wt MoPrP-B (ILn / J) , MoPrP(P101L) [Tg(MoPrP , P101L)196 / Prnp0 / 0 ] , HuPrP(M129) [Tg (HuPrP , M129)440 / Prnp0 / 0 ] , HuPrP(V129) [Tg(HuPrP , V129)152 / Prnp0 / 0 ] , HuPrP(P102L) [Tg(HuPrP , P102L)7 / Prnp0 / 0 ] , and MHu2M(P102L) [Tg(MHu2M , P102L)69 / Prnp0 / 0 ] were processed
GSS(P102L) prions in human patients and Tg mice
We assessed whether the progressive accumulation of abnormal PrP conformers observed in Tg2866 mice applies to GSS patients and to Tg mice expressing the MHu2M transgene with the corresponding P102L mutation , designated Tg(MHu2M , P102L)69 / Prnp0 / 0 mice
These brain samples displayed increased levels of sHuPrPSc (P102L) compared to untreated samples (Fig. 5A , B , and D)
Aberrant HuPrP(P102L) conformers present in the brains of GSS patients and GSS-inoculated Tg mice (A to D) are indistinguishable from those that progressively accumulate in spontaneously ill chimeric Tg(MHu2M , P102L) mice (E to H)
(A to D) Brain homogenates from spontaneously ill Tg(MHu2M , P102L)69 / Prnp0 / 0 mice (lane 2) , Tg(MHu2M , P102L) mice inoculated with homogenates from GSS patients expressing either Val (lane 3) or Met (lane 4) at codon 129 , and a GSS(P102L , M129) patient (lane 5)
Since GSS(P102L) is the only inherited prion disease that has been successfully modeled in mice with respect to generating infectivity de novo , we chose to exploit the P102L mutation
The neuropathologic changes characteristic of humans carrying the GSS(P102L) mutation , including large , multicentric plaques that stain with anti-PrP antibodies , are preserved in Tg(MoPrP , P101L) mice that develop prion disease spontaneously as well as in those that develop disease after inoculation with GSS prions or with the mutant 55-mer -rich peptide
In contrast , when human GSS(P102L) prions are transmitted to nonhuman primates or non-Tg mice , these GSS-type plaques are rarely found (1 , 32 , 46)
Reference #26 (Apetri AC et al.): P102L
All variants containing mutations linked to familial prion diseases (P102L , D178N with 129M and 129V polymorphism , V180I , F198S , E200K , R208H , V210I , and Q217R) were constructed on the background of W99F / Y218W huPrP-(90-231) (21 ) by site-directed mutagenesis using appropriate primers and the QuikChange kit (Stratagene)
These data show that whereas some of the disease-associated mutations produce a pronounced decrease in the global stability of Y218W / W99F huPrP-(90-231) , the effect of others (e.g. P102L and E200K) is essentially negligible
Two notable exceptions are the E200K and P102L variants , in which cases mutations have very little effect on either the global protein stability (as measured by (image)) or the stability of the folding intermediate
Data of Table II indicate that at least for two PrP variants , P102L and E200K , the population of an intermediate is similar to that for the wild-type prion protein , even though the intermediate state is always much more populated than the fully unfolded state
As far as the P102L mutation is concerned , the lack of significant changes in the energy landscape is not surprising because residue 102 maps to a region outside the folded domain
Reference #27 (Zanusso G et al.): P102L
Previous studies have disclosed that an internal PrP fragment of 7-8 kDa detected in patients with P102L Gerstmann-Sträussler-Scheinker disease (7 , 25 ) and a C-terminal fragment associated with familial Creutzfeldt-Jakob disease due to E200K mutation (26 ) are toxic to neurons in combination with or in the absence of PrP expression (27 , 28 )
Reference #28 (Ishida C et al.): P102L
Methods: Eight patients with prion diseases were examined: three with sporadic Creutzfeldt-Jakob disease (sCJD) , two with dural graft associated CJD (dCJD) , one with Gerstmann-Sträussler-Scheinker disease (GSS) with a PrP P102L mutation (GSS102) , and two with a P105L mutation (GSS105)
METHODS (image)TOP (image)ABSTRACT (image)METHODS (image)RESULTS (image)DISCUSSION (image)REFERENCES Patients We studied eight patients with necropsy confirmed prion diseases: three with sCJD , two with dCJD , 11 , 12 one with GSS with a PrP P102L mutation (GSS102) , and two with a PrP P105L mutation (GSS105) (table 1(image) )
(C) Patient 6 with Gerstmann-Sträussler-Scheinker disease (GSS) having a P102L mutation showed both granular PrP deposits and a few PrP plaques
Yamada M , Tomimitsu H , Yokota Y , et al. Involvement of the spinal posterior horn in Gerstmann-Sträussler-Scheinker disease (PrP P102L)
Reference #29 (Tateishi J et al.): P102L
Hereditary cases with P102L , P105L , A117V , Y145stop , and insertions had different features but all demonstrated a long clinical duration and the presence of PrP plaques
The experimental transmission to mice of these mutant forms was difficult , except for one-third of the cases with P102L
Reference #30 (Young K et al.): P102L
The most common mutation causing Gerstmann-Straussler-Scheinker (GSS) disease is P102L in the prion protein
We describe a patient with GSS disease in whom the P102L mutation is in coupling with valine at residue 129
The clinical presentation in P102L-V129 differs greatly from that seen in P102-M129 patients.
Reference #31 (Tanaka Y et al.): P102L
OBJECTIVE: A new variant of Gerstmann-Straussler-Scheinker disease (GSS) was reported , which had a substitution of glutamate to lysine at codon 219 (E219K) in addition to a P102L mutation on the same allele of the PrP gene
CONCLUSION: Neurological and neuropathological features in the patients were atypical of the classic form of GSS with P102L mutation
Reference #32 (Singh N et al.): P102L
Singh Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L J
Reference #33 (Hegde RS et al.): P102L
Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L
Different patterns of truncated prion protein fragments correlate with distinct phenotypes in P102L Gerstmann-Straussler-Scheinker disease
Reference #34 (Parchi P et al.): P102L
95 , Issue 14 , 8322-8327 , July 7 , 1998 Neurobiology Different patterns of truncated prion protein fragments correlate with distinct phenotypes in P102L Gerstmann-Sträussler-Scheinker disease Piero Parchi * , dagger , Shu G
Carleton Gajdusek , May 13 , 1998 ABSTRACT (image) Top (image) Abstract (image) Introduction (image) Materials & Methods (image) Results (image) Discussion The clinicopathological phenotype of the Gerstmann-Sträussler-Scheinker disease (GSS) variant linked to the codon 102 mutation in the prion protein (PrP) gene (GSS P102L) shows a high heterogeneity
Immunoblot analysis of brain homogenates from GSS P102L patients showed two major protease-resistant PrP fragments (PrP-res) with molecular masses of approx 21 and 8 kDa , respectively
To contribute to the definition of the full spectrum of PrP-res types that forms in human prion diseases , we examined the physicochemical properties of the PrP-res fragments extracted from the affected brain of subjects carrying the P102L mutation , the most common mutation linked to GSS
The study of the PrP-res and its correlation with the phenotype of the disease is particularly appropriate in the P102L GSS subtype because this disease shows an heterogeneous phenotype that is not fully explained by variation in the PRNP genotype (8 , 9 )
We show that , in subjects carrying the P102L mutation who were syngenic at PRNP codon 129 and 219 , distinct pathological features of the disease correlate with different patterns of PrP-res fragments
Summary of genetic , clinical , and histopathologic features in the seven P102L GSS patients Tissues
The presence of the P102L mutation was confirmed in all patients by digestion of amplified DNA with the restriction enzyme DdeI
(A) Immunoblot analysis with the 3F4 antibody of the PrP-res extracted from the cerebral cortex of four subjects with GSS P102L (lanes 2-7) and one subjects with sporadic CJD (lane 1)
However , the ratio of the three major glycoforms of PrP-res was significantly different between the GSSP102L samples and sporadic CJD (Fig. 2 )
Relative proportion of the three PrP-res glycoforms of the 21-kDa fragment (PrP-res type 1) in GSS P102L compared with that of sporadic CJD (sCJD)
Immunoblot analysis of purified detergent insoluble fractions (P3) of PrP from the cerebral cortex of one control subject (lane 6) and two subjects (2 and 7) with GSS P102L (lanes 2-5) , before (lanes 1 , 2 , 4 , and 6) and after PK digestion (lanes 3 and 5)
PrP immunoreactivity in two GSS P102L subjects showing distinct patterns of PrP-res on immunoblot
DISCUSSION (image) Top (image) Abstract (image) Introduction (image) Materials & Methods (image) Results (image) Discussion The present findings show that the heterogeneity of the clinicopathologic phenotype associated with the GSS P102L variant is related to the presence and relative abundance of two PrP-res fragments that have a different size and degree of glycosylation
Consistent with this hypothesis is the observation that , among the GSS P102L patients , those showing a CJD-like phenotype have , on average , a significantly shorter course (8 , 9 , 11 )
In GSS P102L (as in the other GSS variants previously studied) , the 8-kDa PrP fragment with ragged N and C termini is consistently found to colocalize with the amyloid plaques and provides a marker for the distinction between CJD and GSS at the molecular level
Our data , which show that , in GSS P102L subjects without significant spongiform degeneration , the 8-kDa fragment is the only detectable PrP-res fragment , underline the importance of the search of smaller PrP-res peptides in addition to PrP 27-30
Spongiform degeneration , although inconspicuous , has been observed in other GSS variants in addition to GSS P102L
The presence of a type 1 PrP-res in GSS P102L and of a type 2 PrP-res in GSS F198S would be consistent with the fact that , as shown for sporadic CJD , valine at codon 129 , which is invariably linked to the F198S mutation , favors the formation of PrP-res type 2 whereas methionine , which cosegregates with the P102L mutation , usually is associated with PrP-res type 1 (5 )
Although all CJD variants and fatal familial insomnia have been propagated successfully in experimental animals , the GSS P102L , as well as an individual GSS case carrying an insertion mutation in PRNP , are the only subtypes of GSS that have been transmitted to date (13 , 26-30 )
The reason why , in a subset of subjects with the P102L mutation , PrP-res type 1 is not detectable is puzzling
Transmission studies , aimed to determine whether brain homogenates from subjects with GSS P102L will induce heterogeneous phenotypes in the recipient syngenic animals , may provide significant clues to answer the question
Similarly , neuropatholopic examination and PrP-res typing of animals inoculated with GSS P102L that developed disease will be important to unravel the critical factors that lead to the formation of the 8-kDa amyloidogenic fragment in GSS P102L
Because all 10 cases of GSS P102L transmitted to date have induced in three different animal species a spongiform encephalopathy indistinguishable from that caused by the inoculation of homogenates from sporadic CJD subject with a typical phenotype (28-30 ) , it seems likely that the formation of amyloid plaques and the 8-kDa PrP-res fragment are specifically linked to the presence of the mutation
Singh Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L J
Reference #35 (Prusiner S et al.): P102L
The P102L mutation was the first PrP mutation to be genetically linked to CNS dysfunction in GSS (Fig. 4 B) (4 ) and has since been found in many GSS families throughout the world (245-247 )
Indeed , a mutation in the protein coding region of the PrP gene has been found in all reported kindred with familial human prion disease ; besides the P102L mutation , genetic linkage has been established for four other mutations (16 , 29-31 )
The P102L mutation of GSS was introduced into the MoPrP transgene , and five lines of Tg(MoPrP-P101L) mice expressing high levels of mutant PrP developed spontaneous CNS degeneration consisting of widespread vacuolation of the neuropil , astrocytic gliosis , and numerous PrP amyloid plaques similar to those seen in the brains of humans who die from GSS(P102L) (248-250 )
Singh Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L J
Indeed , a mutation in the protein coding region of the PrP gene has been found in all reported kindred with familial human prion disease ; besides the P102L muta FIG
Reference #36 (Chiesa R et al.): P102L
Tg(PG14) Mice Are Different from Other Transgenic Models Several different kinds of PrP transgenic mice have been described that spontaneously develop a neurological illness (Westaway et al. 1994(image) express a mutant PrP associated with a human familial prion disease are the P101L mice studied by Prusiner and colleagues that carry the mouse homolog of a GSS-related P102L mutation (Hsiao et al. 1990(image) these mice spontaneously developed neurological symptoms and displayed spongiform degeneration , astrocytic gliosis , and PrP-containing plaques in their brains , as well as a peripheral neuromyopathy not commonly seen in human GSS
2 Later , this classic (ataxic) form of GSS , including the original family described by Gerstmann et al. , 1 was found to be associated with a missense mutation (Pro to Leu) at codon 102 (P102L) of the prion protein (PrP) gene (GSS102)
Gerstmann-Sträussler-Scheinker disease with PRNP P102L mutation and valine at codon 129
Barbanti P , Fabbrini G , Salvatore M , et al. Polymorphism at codon 129 or codon 219 of PRNP and clinical heterogeneity in a previously unreported family with Gerstmann-Sträussler-Scheinker disease (PrP-P102L mutation)
Piccardo P , Ghetti B , Dickson DW , et al. Gerstmann-Sträussler-Scheinker disease (PRNP P102L) : amyloid deposits are best recognized by antibodies directed to epitopes in PrP region 90-165
Singh Cell Surface Accumulation of a Truncated Transmembrane Prion Protein in Gerstmann-Straussler-Scheinker Disease P102L J
Reference #4 (Cardone F et al.): Pro102Leu
We analysed PrP27-30 glycotypes in a large number of TSE-affected patients: 50 sporadic CJD (sCJD) , 1 iatrogenic CJD , 1 Gerstmann-Straussler-Scheinker syndrome (GSS) with the Pro102Leu mutation of PrP , 3 familial CJD (fCJD) with the Glu200Lys mutation and , for the first time , 7 fCJD with the Val210ll3e mutation
Reference #8 (Petraroli R et al.): Pro102Leu
The goodness of this method is demonstrated in the analysis of three sporadic CJD patients with different genotypes at codon 129 and three inherited cases bearing different point mutations of PRNP: the Pro102Leu mutation linked to Gerstmann-Straussler-Scheinker-syndrome , the Val210Ile mutation and a novel mutation at codon 211 (Gln211Glu) both associated to familial CJD.
2005, PrionDB.
cmbi.ru.nl), 22-Aug-2005