PrionDB: Extraction of mutation data from the literature

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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 210 were found are listed after the table.

Point mutations at position V210 in PRIO_HUMAN

ProteinPRIO_HUMAN (P04156)    Gene: PRNP    (other point mutations)Swiss-Prot
Cross-reference table
Family page
PositionV210
General numbering (PrionDB) -
DomainNot determined
Family alignments Mammalian prion proteins
Prion proteins (PRP, PRNP)
Other point mutations at the same position Position 210 in Mammalian prion proteins family
Position 210 in Prion proteins (PRP, PRNP) family
Reference #1Vetrugno V, Malchow M, Liu Q, Marziali G, Battistini A, Pocchiari M
Neurosci Lett 1999 Jul 23;270(1):41-4.		Medline
Text sourceabstract
Point mutationV210I (True positive)
Reference #2Cardone F, Liu QG, Petraroli R, Ladogana A, D'Alessandro M, Arpino C, Di Bari M, Macchi G, Pocchiari M
Brain Res Bull 1999 Aug;49(6):429-33.		Medline
Text sourceabstract
Point mutationV210I (True positive)
Cited point mutationVal210lle
Reference #3Mouillet-Richard S, Teil C, Lenne M, Hugon S, Taleb O, Laplanche JL
J Neurol Sci 1999 Oct 15;168(2):141-4.		Medline
Text sourceabstract
Point mutationV210I (True positive)
Reference #4Zuegg J, Gready JE
Biochemistry 1999 Oct 19;38(42):13862-76.		Medline
Text sourceHTML and PDF full texts
Point mutationV210I (True positive)
Reference #5Petraroli R, Vaccari G, Pocchiari M
J Neurosci Methods 2000 Jun 30;99(1-2):59-63.		Medline
Text sourceabstract
Point mutationV210I (True positive)
Cited point mutationVal210Ile
Reference #6Tagliavini F, Lievens PM, Tranchant C, Warter JM, Mohr M, Giaccone G, Perini F, Rossi G, Salmona M, Piccardo P, Ghetti B, Beavis RC, Bugiani O, Frangione B, Prelli F
J Biol Chem 2001 Feb 23;276(8):6009-15.		Medline
Text sourceHTML full text
Point mutationV210I (True positive)
Reference #7Thompson AJ, Barnham KJ, Norton RS, Barrow CJ
Biochim Biophys Acta 2001 Jan 12;1544(1-2):242-54.		Medline
Text sourceHTML full text
Point mutationV210I (Not yet checked)
Cited point mutationVal210Ile,Val-210-Ile
Reference #8Mastrianni JA, Capellari S, Telling GC, Han D, Bosque P, Prusiner SB, DeArmond SJ
Neurology 2001 Dec 26;57(12):2198-205.		Medline
Text sourceHTML full text
Point mutationV210I (True positive)
Reference #9Capellari S, Parchi P, Wolff BD, Campbell J, Atkinson R, Posey DM, Petersen RB, Gambetti P
Neurology 2002 Nov 26;59(10):1628-30.		Medline
Text sourceHTML full text
Point mutationV210I (True positive)
Reference #10Mastrangelo P, Serpell L, Dafforn T, Lesk A, Fraser P, Westaway D
FEBS Lett 2002 Dec 4;532(1-2):21-6.		Medline
Text sourceHTML and PDF full texts
Point mutationV210I (True positive)
Reference #11Sauer H, Wefer K, Vetrugno V, Pocchiari M, Gissel C, Sachinidis A, Hescheler J, Wartenberg M
Free Radic Biol Med 2003 Sep 15;35(6):586-94.		Medline
Text sourceHTML full text
Point mutationV210I (Not yet checked)
Reference #12Gambetti P, Kong Q, Zou W, Parchi P, Chen SG
Br Med Bull 2003;66:213-39.		Medline
Text sourceHTML full text
Point mutationV210I (True positive)
Reference #13Apetri AC, Surewicz K, Surewicz WK
J Biol Chem 2004 Apr 23;279(17):18008-14. Epub 2004 Feb 2.		Medline
Text sourceHTML full text
Point mutationV210I (True positive)
Reference #14Pocchiari M, Puopolo M, Croes EA, Budka H, Gelpi E, Collins S, Lewis V, Sutcliffe T, Guilivi A, Delasnerie-Laupretre N, Brandel JP, Alperovitch A, Zerr I, Poser S, Kretzschmar HA, Ladogana A, Rietvald I, Mitrova E, Martinez-Martin P, de Pedro-Cuesta J, Glatzel M, Aguzzi A, Cooper S, Mackenzie J, van Duijn CM, Will RG
Brain 2004 Oct;127(Pt 10):2348-59. Epub 2004 Sep 10.		Medline
Text sourceHTML full text
Point mutationV210I (Not yet checked)
Reference #15Ladogana A, Puopolo M, Poleggi A, Almonti S, Mellina V, Equestre M, Pocchiari M
Neurology 2005 May 10;64(9):1592-7.		Medline
Text sourceabstract
Point mutationV210I (Not yet checked)
Reference #16Riek R, Wider G, Billeter M, Hornemann S, Glockshuber R, Wuthrich K
Proc Natl Acad Sci U S A 1998 Sep 29;95(20):11667-72.		Medline
Text sourceHTML and PDF full texts
Point mutationV210I (True positive)

Relevant sentences

Reference #1 (Vetrugno V et al.): V210I
  • We have established stably transfected human neuroblastoma cells (SH-SY5Y) expressing mutant V210I , or wild-type PrPc

Reference #3 (Mouillet-Richard S et al.): V210I
  • A point mutation at codon 210 of the prion protein gene (PRNP) , resulting in the substitution of isoleucine for valine (V210I) has been found in a 54-year-old Moroccan patient affected with Creutzfeldt-Jakob disease (CJD)

  • This patient is the first carrier of the PRNP V210I mutation reported from North Africa

  • The mother of the proband , aged 72 , is a further example of an asymptomatic elderly carrier of the PRNP V210I mutation , suggesting an incomplete penetrance of the disease.

Reference #4 (Zuegg J et al.): V210I
  • 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 #6 (Tagliavini F et al.): V210I
  • 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 )

Reference #7 (Thompson AJ et al.): V210I
  • Peptide synthesis , purification and characterisation Peptides 1 (H3: PrP198– ; 218 NH2-FTETDVKMMERVVEQMCITQY-CO2H ; Mr = 2582) and 2 (H3-V210I: PrP198– ; 218 NH2-FTETDVKMMERVIEQMCITQY-CO2H ; Mr = 2596) were synthesised manually in the solid phase on Wang resin using Fmoc chemistry

Reference #8 (Mastrianni JA et al.): V210I
  • Alert me when: new articles cite this article Download to Citation Manager Collections under which this article appears: Prion Neurology 2001 ; 57:2198-2205 © 2001 American Academy of Neurology ------------------------------------------------------------------------ Articles Inherited prion disease caused by the V210I mutation Transmission to transgenic mice J

  • Methods: The clinicopathologic features of four individuals from the United States who died of fCJD(V210I) were compared

  • Transgenic (Tg) mice expressing a chimeric human-mouse PrP transgene were inoculated with brain extracts from three fCJD(V210I) cases , sporadic CJD (sCJD) , fCJD(E200K) , and fatal familial insomnia (FFI) , to compare prion strain characteristics

  • Results: The clinicopathologic profile of fCJD(V210I) was variable among cases but shared similarities with sCJD

  • 10 The mutation results in the substitution of isoleucine at position 210 for valine (V210I)

  • Transmission of familial CJD (fCJD)(V210I) has not been reported

  • We identified four patients with fCJD(V210I)

  • Brain extracts from three cases of fCJD(V210I) inoculated into transgenic (Tg) mice harboring only the chimeric mouse-human-mouse PrP (Tg[MHu2M]Prnp 0 / 0) produced disease in all mice about 200 days after inoculation

  • Our findings suggest that fCJD(V210I) shares a clinicopathologic and experimental phenotype with sporadic CJD (sCJD) but not with fCJD(E200K) or FFI(D178N , M129)

  • Summary of clinical phenotypes of V210I Patient B

  • Frozen samples from three of the four fCJD(V210I) cases were available for transmission studies , and at least eight Tg(MHu2M)Prnp 0 / 0 mice were inoculated per group

  • Sequence analysis of the entire ORF of the PRNP gene of these patients revealed a single G-to-A substitution at the first nucleotide of codon 210 (see figure 1 ) , resulting in a Val-to-Ile change in the protein (V210I)

  • As with the V210I cases , these were similarly Met homozygous at codon 129

  • Western immunoblot analysis of a sample of frontal cortex from each brain of the three available V210I carriers demonstrated the typical protease-resistant PrP 27 to 30 fragment seen after limited proteolytic digestion

  • The proportion of di- , mono- and unglycosylated PrP Sc and the approximately 21-kD unglycosylated fraction in each V210I case were consistent with the type 1 pattern of PrP Sc seen in sCJD(M / M129) 20 , 21 (figure 2 )

  • Western blot analysis of protease-resistant prion protein (PrP) from a case of sporadic Creutzfeldt-Jakob disease (sCJD) and each of three V210I cases (Patients A , B , and C)

  • Type 1 PrP Sc , defined by a 21-kD relative molecular mass of the unglycosylated form , is the most common PrP Sc type in sCJD (M / M129) and was present in all three V210I cases

  • These findings suggest that the PRNP V210I mutation can cause aberrant trafficking of NFP in neurons and that this abnormality is followed by swelling and achromasia and then by nerve cell death

  • Pathologic profile of prion disease associated with thePRNP-V210I mutation View larger version (136K): [in this window] [in a new window]   Figure 3

  • The neuroanatomic pattern of PrP Sc in Tg mice was the same with each of the three fCJD(V210I) inocula (data not shown)

  • The pattern of PrP Sc deposition with fCJD(V210I) prions was very similar to that associated with sCJD prions , with only slight differences noted (see figure 4 , A and C ) , including lower-intensity , but diffuse , PrP Sc immunostaining throughout the brainstem (see figure 4 , B and D )

  • Differences were more obvious between fCJD(V210I) and fCJD(E200K)

  • 6 These differences in histoblot patterns among the familial prion diseases , along with the atypical pathologic features of fCJD(V210I) , argue that the V210I mutation produces a strain of prions that is similar to those of sCJD but different from that of fCJD(E200K) and FFI

  • Histoblot of brain tissue from Tg(MHu2M)Prnp 0 / 0 mice inoculated with human brain homogenates from cases of familial Creutzfeldt-Jakob disease (fCJD(V210I)) (A , B) , sporadic CJD (sCJD) (C , D) , fCJD(E200K) (E , F) , and fatal familial insomnia (G , H)

  • (image)References In this study , we provide the most complete clinical and neuropathologic analysis of fCJD(V210I) to date

  • This variation in features is consistent with previous reports of the V210I pedigree , which described a 'stroke-like' onset in some and a cerebellar syndrome in other affected patients

  • Prion strains and fCJD(V210I)

  • We found that the PrP Sc accumulation patterns following intracerebral inoculation with brain extracts from three of the fCJD(V210I) cases were very similar , even though the clinical presentation in the four patients with fCJD(V210I) was quite variable

  • First , the V210I mutation in the PRNP gene is linked to a neurodegenerative disorder with the clinical and neuropathologic features of CJD in which infectious prions are formed , verifying that it causes a transmissible spongiform encephalopathy

  • Second , the disease phenotype caused by the fCJD(V210I) prions is very similar to that caused by sCJD prions

  • Third , the accumulation of PrP Sc caused by fCJD(V210I) prions is different from those caused by fCJD(E200K) and FFI(D178N , M129) prions

  • Frequency and penetrance of fCJD(V210I)

  • Based on our findings and those of others , the V210I mutation may be under-represented in some older reports of CJD cases

  • The observed variability in symptom onset and neuropathologic changes that we noted among the V210I patients compares with that reported with fCJD(E200K)

  • The same is likely to be true with the V210I mutation , although the number of cases is currently too small to make such predictions

Reference #9 (Capellari S et al.): V210I
  • In both patients the disease duration exceeds that of familial forms of CJD linked to the E200K or V210I mutations that have phenotypes similar to that of the sporadic CJD MM1 subtypes

Reference #10 (Mastrangelo P et al.): V210I
  • Properties of the V180I and V210I F-CJD mutations 3.5

  • On the other hand , a relationship was noted for five F-CJD mutations (V180I , E196K , E200K , V210I , and E211Q: Table 1 )

  • Of these , V180I , E196K , E200K and E211Q result in amino acid changes that are exactly conserved in human Dpl , while V210I involves an isoleucine conserved in mouse , cow , sheep and rat Dpl proteins ( Fig. 1 )

  • Properties of the V180I and V210I F-CJD mutations V180I and V210I are conservative changes in residues almost directly opposite one another on the inside faces of α ; B and α ; C within PrP′ ; s hydrophobic core [34 ]

  • When modeled by molecular mechanics on the wt HuPrP NMR these analyses revealed a substantial change in the RMSD values for the V180I (2.26 Å ; ) and V210I (2.13 Å ; ) proteins compared to wt HuPrP (3.5 Å ; ) , an alteration again in the direction of the Dpl structure

  • Considered collectively , averaged RMSD values per atom for V180I , E196K , E200K , V210I and E211Q were 2.24± ; 0.09 Å ; , versus 3.50 and 3.45 Å ; for wt PrP and D178N PrP , respectively

  • The data also showed a tendency for a greater number of superimposed residues and identical residues in the alignment when again considering V180I , E196K , E200K , V210I and E211Q mutant proteins versus wt PrP and D178N PrP (Table 1 )

  • On the other hand , a relationship was noted for ¢ve F-CJD mutations (V180I , E196K , E200K , V210I , and E211Q: Table 1)

  • Wild type 61 3.50 13 D178N (11) G(S) A , E , G , H , Y , V 178N 61 3.45 13 V180I (4) I (I) A , D , G , L , F 180I 59 2.26 12 E196K (3) K(K) A , D , E , G , Q , V , Stop 196K 71 2.38 15 E200K (63) K(K) A , D , E , G , V , Stop 200K 66 2.18 18 200K NMR structure 68 1.87 15 V210I (5) V(I) n.a

  • Properties of the V180I and V210I F-CJD mutations V180I and V210I are conservative changes in residues almost directly opposite one another on the inside faces of KB and KC within PrPPs hydrophobic core [34]

  • When modeled by molecular mechanics on the wt HuPrP NMR these analyses revealed a substantial change in the RMSD values for the V180I (2.26 Aî ) and V210I (2.13 Aî ) proteins compared to wt HuPrP (3.5 Aî ) , an alteration again in the direction of the Dpl structure

  • Considered collectively , averaged RMSD values per atom for V180I , E196K , E200K , V210I and E211Q were 2.24 þ 0.09 Aî , versus 3.50 and 3.45 Aî for wt PrP and D178N PrP , respectively

Reference #11 (Sauer H et al.): V210I
  • The correlation of PrPc expression to the intracellular ROS levels was investigated by the use of neuroblastoma cells overexpressing either mutant V210I PrP , or wild-type PrPc

  • It was observed that the intracellular redox state was significantly reduced in PrPc as well as V210I PrP overexpressing cells as compared to non-transfected cells

  • Author Keywords: Prion ; Multicellular tumor spheroid ; Reactive oxygen species ; Growth factors ; Tumor necrosis factor- alpha ; NADPH-oxidase , Free radicals Article Outline • ; Introduction • ; Materials and methods • ; Culture technique of multicellular spheroids • ; Incubation of Gli-36 tumor spheroids with growth factors and TNF- alpha • ; Transfection of SH-SY5Y cells • ; Im munohistochemical techniques and semiquantitative im munohistochemistry • ; Enzymatic dissociation of Gli-36 tumor spheroids • ; Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) • ; Determination of intracellular ROS levels • ; Statistical analysis • ; Results • ; Elevation of intracellular ROS levels upon incubation of multicellular Gli-36 glioma tumor spheroids with growth factors and cytokines • ; Elevation of PrP c following treatment with growth factors and TNF- alpha ; effects of antioxidants and role of NADPH-oxidase • ; Effects of growth factors and TNF- alpha on PrP gene expression • ; Effects of PrP c and V210I PrP overexpression on intracellular ROS levels and ATP-induced ROS generation • ; Discussion Introduction The cellular form of the prion protein (PrPc) is a highly conserved cell surface glycoprotein that is expressed predominantly in neuronal cells but also in a variety of other cell types

  • By the use of neuroblastoma tumor spheroids it is demonstrated that overexpression of either wild type PrPc or mutant V210I PrP resulted in significant reduction of intracellular ROS levels and decreased growth factor-induced ROS generation , which clearly indicates that PrP plays a role in the cellular antioxidant system

  • In brief , SH-SY5Y were transfected with wt or mutant (V210I) PrP by calcium phosphate DNA coprecipitation (Invitrogen , Karlsruhe , Germany) , and selected in G418 (Gibco)

  • Human PRP ORFs , derived from a normal individual (wt) and V210I familial CJD (V210I) patient were amplified by PCR and cloned into the eukaryotic expression vector pRcCMV (Invitrogen)

  • Effects of PrP c and V210I PrP overexpression on intracellular ROS levels and ATP-induced ROS generation In the present study it is hypothesized that endogenous PrP c is participating in the intracellular antioxidative defense

  • To investigate this issue , multicellular tumor spheroids were grown from transfected cell lines of SH-SY5Y neuroblastoma cells , which overexpressed either wt PrP c or mutant V210I PrP

  • Patients carrying the V210I mutation show clinical , neuropathological , and biochemical features nearly indistinguishable from those of sporadic CJDs [19 and 20 ]

  • It was shown that in SH-SY5Y tumor spheroids overexpressing either wt PrP c or mutant V210I PrP displayed significant decreased intracellular ROS levels as compared to control cells and cells transfected with empty vector (Fig. 6 )

  • Furthermore , it was clearly shown that the elevation of ROS following sti mulation of cells for 90 min with ATP was completely abolished in wt PrP c and mutant V210I PrP overexpressing cell lines , which clearly indicates that PrP may be involved in the cellular antioxidative defense

  • Intracellular ROS levels and ATP-induced ROS generation in neuroblastoma SH-SY5Y tumor spheroids overexpressing either wt PrP c (SH-SY5Y , wt PrP c) or V210I PrP (SH-SY5Y , V210I PrP)

  • The notion of a physiological role of PrP c in the cellular antioxidative defense was validated by the use of cell lines overexpressing either wt PrP c , or mutant V210I PrP , which carries a point mutation in the PrP gene resulting in a sporadic CJD phenotype [19 and 20 ]

  • It has been previously shown that unlike PrP Sc , V210I PrP and PrP c are located in cell membranes and are susceptible to proteinase K digestion [16 ]

  • Our data clearly demonstrate that intracellular ROS levels were significantly decreased in PrP c as well as V210I PrP overexpressing cell lines , which strongly indicates an antioxidant function

  • Moreover , it was evidenced that the increase in ROS generation following sti mulation of cells with ATP was totally abolished , which suggests that overexpression of PrP c as well as V210I PRP protein may result in a loss of growth factor-induced cell signaling function because ROS are involved in the signal transduction cascade elicited following purinergic receptor sti mulation [18 ]

Reference #12 (Gambetti P et al.): V210I
  • Lane 1 , sCJD 129MM , PrPSc type 1 ; lane 2 , sCJD 129VV , PrPSc type 2 ; lane 3 , fCJD E200K-129M , PrPSc type 1 ; lane 4 , fCJD E200K-129V , PrPSc type 2 ; lane 5: fCJD 144bp-Insert-129M , PrPSc type 1 ; lane 6 , fCJD V210I-129M , PrPSc type 1

  • CJD with the V210I-129M haplotype (CJD V210I-129M) The V210I-129M haplotype has been reported in 23 subjects52

  • The V210I mutation exhibits low penetrance

  • Histopathological features CJDV210I-129M is characterised by spongiosis and gliosis of the grey matter , often more prominent in the cerebral cortex and molecular layer of the cerebellum81 , 82 , 84

  • The V210I PrPSc is of type 16 , 84 , 85 and derived from both the mutant and normal PrP77 , 86

  • CJDV210I-129M has been transmitted to humanised transgenic mice90 that showed a PrPSc distribution similar to that produced by sCJD and different from that of other familial prion diseases84

  • Mastrianni JA , Capellari S , Telling GC et al. Inherited prion disease caused by the V210I mutation: transmission to transgenic mice

Reference #13 (Apetri AC et al.): V210I
  • 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)

  • Fig. 3 shows representative kinetic traces for the refolding and unfolding of the wild-type huPrP-(90-231) and the V210I variant

  • Representative kinetic traces for the refolding (left panels) and unfolding (right panels) of the wild-type huPrP-(90-231) and the V210I variant

  • The results of such experiments for two huPrP-(90-231) variants , F198S and V210I , are shown in Fig. 5

  • The effect of protein concentration on refolding kinetics of huPrP-(90-231) variants F198S and V210I

  • Representative kinetic traces for the refolding of V210I huPrP-(90-231) ({circ}) together with control stopped-flow data for NATA ()

  • For example , the V210I mutation results in only a minor decrease in (image) , whereas the intermediate for this protein is one of the most stable among all PrP variants tested

  • The reaction coordinate diagram for the V210I variant , together with that for the wild-type PrP , is shown schematically in Fig. 7 View larger version (16K): [in this window] [in a new window]   FIG

  • The reaction coordinate diagram for the refolding of the wild-type huPrP-(90-231) (red) and the V210I variant (blue)

  • The (image) value for the V210I variant is lower than that for the wild-type huPrP-(90-231) , indicating higher population of the intermediate for the mutant protein

Reference #14 (Pocchiari M et al.): V210I
  • The Bonferroni correction for multiple testing was adopted within the five subgroups of gTSE (E200K and V210I gCJD , GSS , FFI and insert mutations) and the two subgroups of iCJD patients

  • An analysis was only performed for the two most frequent PRNP mutations (i.e. E200K and V210I)

  • Median clinical duration (Table 5 ) for E200Kand V210I patients was 4 months

  • In the univariate Cox analysis , survival of both E200K and V210I patients was shorter in males than in females

  • Though both mutations co-segregate with methionine at codon 129 , methionine homozygosity was associated with a shorter survival in E200K patients , but a long survival in V210I patients (Table 5 , Fig. 3 )

  • Survival was significantly shorter in patients with late age at onset only in V210I patients (Table 5 )

  • Multivariate analyses confirmed the significance of all variables for V210I patients (Table 5 )

  • View this table: [in this window] [in a new window]   Table 4 Clinical duration of disease in genetic CJD patients carrying different point mutations of the PRNP gene View this table: [in this window] [in a new window]   Table 5 Survival times for genetic CJD (E200K and V210I) View larger version (29K): [in this window] [in a new window]   Fig. 3 Influence of codon 129 genotype on Kaplan-Meier survival curves in (A) E200K CJD , (B) V210I CJD , (C) FFI , (D) human growth hormone-related CJD

Reference #15 (Ladogana A et al.): V210I
  • The most frequent mutation was the V210I (n = 54) , and the second most common the E200K (n = 42)

Reference #16 (Riek R et al.): V210I
  • Close-ups of individual mutation sites: (a) D178N and T183A , (b) Q217R , (c) F198S , and (d) V180I and V210I

Reference #7 (Thompson AJ et al.): Val-210-Ile
  • The Val-210-Ile pathogenic Creutzfeldt– ; Jakob disease mutation increases both the helical and aggregation propensities of a sequence corresponding to helix-3 of PrP C Andrew J

  • CD and NMR data indicate that helical interactions , stabilised by the Val-210-Ile mutation , may precede the formation of small beta , Greek-sheet aggregates in this peptide sequence

  • To investigate the influence of the familial Val-210-Ile mutation on local helical stability we have synthesised two peptides corresponding to helix-3 with and without the pathogenic mutation (peptides 1 and 2 in Fig. 1 )

  • Results are interpreted in relation to the influence of the Val-210-Ile mutation on the local sequence within full length PrP

  • Peptide 2 , corresponding to the helical region 3 of PrP C with the pathological Val-210-Ile mutation , is also random coil at pH 6.0 and above

  • Thermal stability of peptides 1 and 2 To determine the influence of the PrP Val-210-Ile mutation on helical stability , the helical stabilities of peptides 1 and 2 were determined by comparison of their thermal denaturation profiles using CD spectroscopy

  • The slopes of the curves are similar , indicating that the Val-210-Ile mutation increases the small alpha , Greek-helical propensity of this sequence , but does not stabilise the helix to thermal denaturation

  • Aggregation propensity of peptides 1 and 2 To determine if the Val-210-Ile mutation influences the aggregation propensity of the PrP C helix-3 sequence , the time-dependent turbidities of peptides 1 and 2 were compared

  • Influence of the valine-210-isoleucine mutation on local helical stability Structural , conformational and aggregation studies of two peptides corresponding to helical region 3 of PrP C , one corresponding to the native sequence and the other containing the Val-210-Ile mutation , indicate that this mutation increases both the small alpha , Greek-helical and aggregation propensities of this sequence

  • The Val-210-Ile mutation does not stabilise the helix by a modification of local helical structure , but instead increases the helical propensity of this region

  • The increased helical propensity due to the Val-210-Ile mutation observed in aqueous TFE may be because isoleucine has a slightly higher helical propensity than valine in a membrane-like environment and in soluble proteins [33 and 34 ]

  • In aqueous buffer , peptide 2 also has a higher small alpha , Greek-helical propensity than does peptide 1 , indicating that the Val-210-Ile mutation stabilises small alpha , Greek-helical structure in both hydrophobic and hydrophilic environments

  • The increased aggregation propensity of peptide 2 as compared with peptide 1 indicates that the Val-210-Ile mutation facilitates intermolecular interactions , either by increasing small beta , Greek-sheet propensity or by facilitating dimerisation of helices , with subsequent small beta , Greek-sheet formation

  • What is surprising about the results presented here is that the pathogenic Val-210-Ile mutation initially increases the small alpha , Greek-helical propensity of a peptide corresponding to helix-3 of the prion protein before subsequent conversion to small beta , Greek-sheet

  • The greater helical tendency of the Val-210-Ile mutant can be viewed as circumstantial evidence for the presence of helix– ; helix interactions such as coiled-coils in the reaction pathway

  • Therefore , the increased helical propensity of the Val-210-Ile mutant peptide 2 , as compared with peptide 1 , observed in the hydrophobic solvent TFE / water , indicates that this mutation should also stabilise helix-3 within full length PrP C

  • Our NMR structural comparison of peptides 1 and 2 indicates that the Val-210-Ile mutation does not modify the structure of the helix formed in this region

  • Because the Val-210-Ile mutation increases the aggregation propensity of the helix-3 sequence , it may also preferentially stabilise a PrP isoform with small beta , Greek-sheet rather than small alpha , Greek-helical structure within this region

  • A second peptide corresponding to the same sequence with the pathogenic Val-210-Ile prion mutation formed either pH-dependent small beta , Greek-sheet or small alpha , Greek-helix in aqueous solution , indicating a higher helical propensity of this mutant sequence

  • The amino acid sequences of peptide 1 , corresponding to residues 198– ; 218 of the prion protein (helix-3) , and peptide 2 which incorporates the familial Val-210-Ile mutation

Reference #5 (Petraroli R et al.): Val210Ile
  • 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.

Reference #2 (Cardone F et al.): Val210lle
  • Type 1A is also formed in Val210lle fCJD

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F.Horn (priondbcmbi.ru.nl), 22-Aug-2005