ライフサイエンスコーパス: PrP-res

1 PrP-res conformation differs among TSE agents derived fr

2 PrP-res formation was significantly reduced by deletion

3 PrP-res isolated from the brains of hamsters infected wi

4 PrP-res molecules that accumulate in the brain and lymph

5 PrP-res patterns in FU- and SY-infected GT1 cells were i

6 PrP-res was also a poor predictor of infectivity because

7 dicate that the conformations of HY and 263K PrP-res differ from DY PrP-res at least in structural re

8 Infrared spectra of HY and 263K PrP-res each had major absorption bands in the amide I r

9 ice expressing anchorless PrP-sen accumulate PrP-res and replicate infectivity.

10 t infection did not necessarily follow acute PrP-res formation.

11 formational conversion of PrPC to additional PrP-res represents agent replication.

12 (residues 90-140) are essential for adopting PrP-res conformation and demonstrate that methionine oxi

13 ha-helix of PrP-sen can fundamentally affect PrP-res formation and/or PrP-sen processing.

14 P-sen glycosylation can significantly affect PrP-res formation.

15 Glycosylation affected PrP-res formation by influencing the amount of PrP-sen b

16 Although PrP-res has been posited as the infectious agent, purifi

17 exposed to 22L, both anchored and anchorless PrP-res were detected over multiple passes.

18 xpressing anchorless PrP-sen made anchorless PrP-res in the first 96 h postinfection, no PrP-res was

19 elation was observed between infectivity and PrP-res detection.

20 ed a discrepancy between TSE infectivity and PrP-res levels in both natural and experimental cases of

21 Here, the levels of TSE infectivity and PrP-res within a peripheral tissue of this mouse model w

22 Here we report that, if PrP-sen and PrP-res are derived from different species, PrP-sen glyc

23 ermolecular interactions between PrP-sen and PrP-res are required to complete the process of conversi

24 y influenced by homology between PrP-sen and PrP-res at amino acid residue 138, a residue located in

25 acid sequence similarity between PrP-sen and PrP-res influences both PrP-res formation and cross-spec

26 ent conversion reactions between PrP-sen and PrP-res.

27 the sites of interaction between PrP-sen and PrP-res.

28 roteinase K-resistant protein referred to as PrP-res or PrPsc accumulates in brain.

29 Here we show that the CWD-associated PrP-res (PrP(CWD)) of cervids readily induces the conver

30 ed proteolysis to yield resistant PrP bands (PrP-res).

31 Thus, the discrepancy between PrP-res and TSE infectivity was also present in the peri

32 Amino acid sequence homology between PrP-res and PrP-sen is important in the formation of new

33 consequence of a complex interaction between PrP-res, PrP-sen, and the cell and may indicate the cell

34 Sc)) involves selective interactions between PrP-res and its normal protease-sensitive counterpart, P

35 peptides may mimic contact surfaces between PrP-res and PrP-sen and thereby serve as models of poten

36 Only PrP-sen was observed to bind PrP-res.

37 amster PrP (Ha119-136) can selectively block PrP-res formation in cell-free systems and scrapie-infec

38 between PrP-sen and PrP-res influences both PrP-res formation and cross-species transmission of infe

39 oduce retinal degeneration, whereas in brain PrP-res production by neurons or astrocytes alone was su

40 all three lines of mice high levels of brain PrP-res accompanied by neurodegeneration were observed.

41 ersion of human PrP-sen to PrP-res driven by PrP-res associated with both scrapie (PrP[Sc]) and BSE (

42 ity to inhibit the polymerization of PrPC by PrP-res.

43 inase K-resistant PrP(Sc)-like conformation (PrP-res).

44 ion was observed in brain regions containing PrP-res amyloid plaques, and a more dispersed conversion

45 >30% of cells displayed abundant cytoplasmic PrP-res aggregates that may trap agent.

46 cuolar pathology but little or no detectable PrP-res.

47 uct was detected in areas containing diffuse PrP-res deposits.

48 required for both generating and disrupting PrP-res formation.

49 ations of HY and 263K PrP-res differ from DY PrP-res at least in structural regions with beta-sheet s

50 These bands were not evident in the DY PrP-res spectra, which had a unique band at 1629-1630 cm

51 s to additional polymerization of endogenous PrP-res aggregates and is analogous to the process of Pr

52 is the binding of soluble PrPC to endogenous PrP-res deposits.

53 Both strains evoked PrP-res in cultured murine cells, although SY induced Pr

54 form resists conversion induced by exogenous PrP-res.

55 of internalizing and disseminating exogenous PrP-res.

56 Similar trafficking of exogenous PrP-res by cortical neurons cultured from the brains of

57 raction of SCRL-bound PrP-sen with exogenous PrP-res as contained in microsomes.

58 microsomes or purified, detergent-extracted PrP-res.

59 in vivo, suggesting that they may facilitate PrP-res formation.

60 le PrPC into both nonfibrillar and fibrillar PrP-res deposits in TSE-infected brain.

61 -fold less protease-resistant than bona fide PrP-res derived from TSE-infected brain tissue, and they

62 The final PrP-res glycoform pattern, however, can be influenced by

63 , and lymph node were analyzed in detail for PrP-res and infectivity.

64 rapie, suggest that analysis of placenta for PrP-res could be the basis for an antemortem test for sh

65 ves as a critical ligand and/or receptor for PrP-res in the course of PrP-res propagation and pathoge

66 a219-232 epitope itself was not required for PrP-res binding; thus, inhibition by alpha219-232 was li

67 re we show that rabbit PrP-sen does not form PrP-res in murine tissue culture cells persistently infe

68 and they showed no increased ability to form PrP-res in a cell-free system.

69 sen or PrP(C)) to a protease-resistant form (PrP-res or PrP(Sc)) is commonly thought to be required i

70 y folded, partially protease-resistant form (PrP-res) of the normal protease-sensitive prion protein

71 to the abnormal proteinase K-resistant form (PrP-res).

72 in or PrP-sen, to a protease-resistant form (PrP-res).

73 sen, to an abnormal protease-resistant form, PrP-res.

74 en) to the abnormal protease-resistant form, PrP-res.

75 sen) to an abnormal protease-resistant form, PrP-res.

76 The newly formed PrP-res associated with the slices in a pattern that cor

77 two major protease-resistant PrP fragments (PrP-res) with molecular masses of approximately 21 and 8

78 Rather, homology between PrP-sen and hamster PrP-res at amino acid residue 155 determined the efficie

79 rotease-resistant product induced by hamster PrP-res.

80 e reaction composed predominantly of hamster PrP-res and PrP-sen.

81 l species, we assayed the ability of hamster PrP-res to convert a panel of recombinant PrP-sen molecu

82 However, PrP-res patterns sometimes differed among individual ani

83 SN56 cells was accompanied by a decrease in PrP-res formation.

84 f exposure of new protease cleavage sites in PrP-res between residues 130 and 157, suggesting that th

85 etween cells; or (iii) insertion of incoming PrP-res into the raft domains of recipient cells.

86 he binding reaction was tested by incubating PrP-res with cell lysates or culture supernatants.

87 These data indicate PrP-res neither encodes nor alters agent-specific charac

88 n cultured murine cells, although SY induced PrP-res only transiently.

89 slational modifications to PrP-sen influence PrP-res formation in vitro.

90 le most anti-scrapie agent compounds inhibit PrP-res formation in vitro, many PrP-res inhibitors have

91 nous sulfated glycans can profoundly inhibit PrP-res accumulation and serve as prophylactic anti-TSE

92 d, critical amino acid residues that inhibit PrP-res formation are located throughout the rabbit PrP

93 h mouse and hamster PrP, was able to inhibit PrP-res formation in both the mouse and hamster cell-fre

94 Tetrapyrroles also inhibited PrP-res formation in a cell-free reaction composed predo

95 ent of the spice turmeric, potently inhibits PrP-res accumulation in scrapie agent-infected neuroblas

96 of the cell type and scrapie strain, initial PrP-res formation occurred rapidly in cells.

97 When normalized for quantity of input PrP-res, scrapie brain microsomes induced dramatically e

98 Internalized PrP-res was then transported along neurites to points of

99 t scrapie infection, SN56 cells internalized PrP-res aggregates into vesicles positive for markers fo

100 al protease-resistant prion protein isoform (PrP-res).

101 to its abnormal protease-resistant isoform (PrP-res) is a major feature of the pathogenesis associat

102 n (PrP-sen) to a protease-resistant isoform (PrP-res) is an important event in pathogenesis.

103 PrP-sen) and the protease-resistant isoform (PrP-res), a model system was employed using PrP-sen reco

104 (PrP-sen) to the protease-resistant isoform (PrP-res).

105 , aggregated protease-resistant PrP isoform, PrP-res, associated with clinical CJD and other transmis

106 s converted to a protease-resistant isoform, PrP-res, by an apparent self-propagating activity of the

107 rP-sen, into its protease-resistant isoform, PrP-res.

108 al change in PrP-sen that may potentiate its PrP-res-induced conversion.

109 prion protein (PrP-res), fluorescent-labeled PrP-res was used to infect a neuronally derived murine c

110 Because superinfected mice had little PrP-res just before the onset of clinical disease and re

111 f infectivity because SY cells that had lost PrP-res were approximately 10-fold more infectious than

112 brain, the fast CJD strain, FU, elicits many PrP-res deposits, whereas the slow SY strain elicits few

113 nds inhibit PrP-res formation in vitro, many PrP-res inhibitors have no activity in vivo.

114 ed the ability of hamster PrP to block mouse PrP-res formation in scrapie-infected mouse neuroblastom

115 reviously shown that the generation of mouse PrP-res was strongly influenced by homology between PrP-

116 s and activated microglia but had negligible PrP-res (the more protease-resistant form of host PrP) o

117 PrP-sen is important in the formation of new PrP-res and thus in the efficient transmission of infect

118 raise the possibility that generation of new PrP-res during TSE infection requires (i) removal of PrP

119 e from clinically ill mice with little or no PrP-res detectable, similar short incubation periods to

120 PrP-res in the first 96 h postinfection, no PrP-res was detected at later passes.

121 prion protein (PrP-res) or in the absence of PrP-res by detection of infectivity.

122 to residue 113 further reduced the amount of PrP-res formed.

123 sequence of PrP-sen influenced the amount of PrP-res generated in the post-binding conversion step.

124 tion of cells is aided by the association of PrP-res with membranes and/or other microsomal constitue

125 However, the molecular basis of PrP-res glycoform variation between different TSE agents

126 effect of this region on the conformation of PrP-res generated in an in vitro cell-free conversion as

127 and/or receptor for PrP-res in the course of PrP-res propagation and pathogenesis in vivo.

128 with the pre-existing brain distribution of PrP-res.

129 The combined effects of PrP-res production in multiple cell types was required t

130 Because the formation of PrP-res fragments of 7-8 kDa with ragged N and C termini

131 if this region was required for formation of PrP-res in a cell-free assay.

132 t residue 138 also affected the formation of PrP-res in a different animal species, we assayed the ab

133 nts significantly inhibited the formation of PrP-res in Sc(+)-MNB cells and had a greatly reduced abi

134 tive compound, 2a, inhibits the formation of PrP-res in SMB cells with an EC50 of 25-50 nM.

135 e events leading to the initial formation of PrP-res may differ from those required for sustained PrP

136 nt in which the strain-specific formation of PrP-res occurs.

137 ree TSE strains resulted in the formation of PrP-res with different conformations using limited prote

138 n shown to inhibit the in vitro formation of PrP-res, a protease-resistant protein critical for TSE p

139 During the formation of PrP-res, PrP-sen undergoes conformational changes that i

140 P-sen that are important in the formation of PrP-res, the exact role of PrP-sen secondary structures

141 However, sustained generation of PrP-res and persistent infection did not necessarily fol

142 A number of analogues showed inhibition of PrP-res infectivity at nanomolar concentrations.

143 phyrins and phthalocyanines as inhibitors of PrP-res accumulation.

144 dings introduce a new class of inhibitors of PrP-res formation that represents a potential source of

145 id not become scrapie-infected, the level of PrP-res in the 22L-infected cells rapidly increased in t

146 Infected SN56 cells released low levels of PrP-res into the culture supernatant, which also efficie

147 ochemical analysis showed that low levels of PrP-res were present in the spleen tissue in comparison

148 e of a concomitant increase in the number of PrP-res-producing cells.

149 Furthermore, the protein banding pattern of PrP-res in these cells changed over time as the cells be

150 ggregates and is analogous to the process of PrP-res fibril and subfibril growth in vivo.

151 the infectious TSE agent consists solely of PrP-res and that PrP-res-induced conformational conversi

152 es from scrapie-infected mice as a source of PrP-res.

153 The amino acid sequence specificity of PrP-res formation correlates with, and may account for,

154 de evidence that the sequence specificity of PrP-res formation in this model is determined more by th

155 nism controlling the sequence specificity of PrP-res formation, we compared the binding of PrP-sen to

156 e of CJD agent characteristics from those of PrP-res, two different mouse-passaged CJD strains were p

157 rion diseases largely depends on the type of PrP-res fragment that forms in vivo.

158 Thus, by its effects on PrP-res conformation, the flexible N-terminal region of

159 y paradoxical effects of sulfated glycans on PrP-res formation, we have assayed their direct effects

160 Persistent PrP-res formation and scrapie infection was restricted t

161 tain cells, we followed acute and persistent PrP-res formation upon exposure of cells to different sc

162 mes induced dramatically enhanced persistent PrP-res formation compared to purified PrP-res.

163 Cells expressing anchored PrP-sen produced PrP-res after exposure to 22L.

164 long-standing practical problem in producing PrP-res fibrils from full-length PrP, and may help in id

165 rmation of protease-resistant prion protein (PrP-res or PrP(Sc)) involves selective interactions betw

166 g abnormal protease-resistant prion protein (PrP-res) formation in scrapie agent-infected cells, we t

167 ssociated, protease-resistant prion protein (PrP-res) from the normal protease-sensitive isoform (PrP

168 ulation of protease-resistant prion protein (PrP-res) is a prime strategy in the development of poten

169 f abnormal protease-resistant prion protein (PrP-res) or in the absence of PrP-res by detection of in

170 nfectious, protease-resistant prion protein (PrP-res), fluorescent-labeled PrP-res was used to infect

171 l, protease-resistant form of prion protein (PrP-res).

172 ivity of the abnormal form of prion protein (PrP-res).

173 neuropathology or pathologic prion protein (PrP-res).

174 d the abnormal isoform of the prion protein, PrP-res, and displayed spongiform degeneration.

175 egated and protease-resistant prion protein, PrP-res, from a normally soluble, protease-sensitive and

176 The effect of disease-associated PrP (PrP-res) association with microsomal membranes on infect

177 hey convert PrP into protease-resistant PrP (PrP-res) but also exert protective activity.

178 imately 10-fold more protease-resistant PrP (PrP-res) in FU brains during terminal disease.

179 iency with which the protease-resistant PrP (PrP-res) of one species induces the in vitro conversion

180 d scrapie-associated protease-resistant PrP (PrP-res) were detected in retina and brain before clinic

181 orm of the host-encoded prion protein (PrP), PrP-res.

182 -associated isoform (protease-resistant PrP; PrP-res) appears to be primarily restricted to cells of

183 en posited as the infectious agent, purified PrP-res itself is not infectious.

184 stent PrP-res formation compared to purified PrP-res.

185 t and the conformation of protease-resistant PrP-res produced from N-terminally truncated PrP-sen.

186 Retinal PrP-res was present in high amounts in both tg7 and tgNS

187 ence of proteinase K (PK)-resistant PrP(Sc) (PrP-res) in postmortem tissues as an indication of TSE d

188 icroglia and astrocytes preceded significant PrP-res accumulation by more than 50 days.

189 In some species, PrP-res accumulates in other tissues as well.

190 support the hypothesis that strain-specific PrP-res conformers can self-propagate by converting the

191 t PrP-res itself can dictate strain-specific PrP-res glycoforms.

192 Thus, strain-specific PrP-res glycosylation profiles are likely the consequenc

193 sulfate and pentosan polysulfate stimulated PrP-res formation.

194 ficiency was examined by comparing sustained PrP-res production in cells treated with either scrapie

195 may differ from those required for sustained PrP-res formation and infection.

196 s, scrapie microsomes induced less long-term PrP-res production than suspended microsomes.

197 e approximately 10-fold more infectious than PrP-res-positive cultures.

198 SE agent consists solely of PrP-res and that PrP-res-induced conformational conversion of PrPC to add

199 These studies provide direct evidence that PrP-res formation involves the incorporation of soluble

200 Y and FU at terminal stages, indicating that PrP-res content does not correlate with infectivity.

201 Here we report that PrP-res itself can dictate strain-specific PrP-res glyco

202 mortem test for sheep scrapie, and show that PrP-res, scrapie infectivity, and scrapie disease are cl

203 Thus, our data suggest that PrP-res molecules isolated from scrapie-infected brains

204 ous recombinant PrP fibrils, suggesting that PrP-res is internalized by a relatively nonspecific pino

205 00-223 (Ha200-223) also potently inhibit the PrP-res induced cell-free conversion of PrP-sen to the p

206 ed by alterations in the conformation of the PrP-res generated.

207 Analysis of the relative amounts of the PrP-res glycoforms has been used to discriminate TSE str

208 owever, after limited digestion with PK, the PrP-res from the DY strain exhibited a fragmentation pat

209 The 21-kDa fragment, similar to the PrP-res type 1 described in Creutzfeldt-Jakob disease, w

210 Thus, PrP-res formation in transgenic mice expressing anchorle

211 showed that heterologous PrP-sen can bind to PrP-res with little conversion to the protease-resistant

212 This highly selective binding to PrP-res and the localized nature of the binding site on

213 nding to PrP-sen and blocking its binding to PrP-res.

214 y influencing the amount of PrP-sen bound to PrP-res, while the amino acid sequence of PrP-sen influe

215 whether these mutants could be converted to PrP-res in both scrapie-infected neuroblastoma cells (Sc

216 RL-bound GPI(+) PrP-sen was not converted to PrP-res until PI-PLC was added to the reaction or the co

217 DRM-associated PrP-sen was not converted to PrP-res until the PrP-sen was either released from DRMs

218 , SCRL-bound GPI(-) PrP-sen was converted to PrP-res without PI-PLC or PEG treatment.

219 bridges stabilize PrP-sen from converting to PrP-res.

220 inhibits the cell-free conversion of PrP to PrP-res.

221 itu conversion of protease-sensitive PrPC to PrP-res in TSE-infected brain slices.

222 219-232 inhibited the binding of PrP-sen to PrP-res and the subsequent generation of PK-resistant Pr

223 found limited conversion of human PrP-sen to PrP-res driven by PrP-res associated with both scrapie (

224 the conformational transition of PrP-sen to PrP-res has not yet been defined.

225 ation, we compared the binding of PrP-sen to PrP-res with its subsequent acquisition of protease resi

226 fically inhibit the conversion of PrP-sen to PrP-res without apparent cytotoxic effects.

227 ot the binding, of the homologous PrP-sen to PrP-res.

228 e important for the conversion of PrP-sen to PrP-res.

229 phalopathies is the conversion of PrP-sen to PrP-res.

230 In this study, we used PrP-res derived from animals infected with two different

231 rmore, FU titers increased 650-fold, whereas PrP-res remained constant.

232 were propagated in neuronal cell lines whose PrP-res patterns differ markedly from each other and fro

233 fated glycosaminoglycans are associated with PrP-res deposits in vivo, suggesting that they may facil

234 direct interactions of the tetrapyrrole with PrP-res and/or PrP-sen.