TY - JOUR
T1 - Further characterization of glycoform-selective prions of variably protease-sensitive prionopathy
AU - Zhang, Weiguanliu
AU - Xiao, Xiangzhu
AU - Ding, Mingxuan
AU - Yuan, Jue
AU - Foutz, Aaron
AU - Moudjou, Mohammed
AU - Kitamoto, Tetsuyuki
AU - Langeveld, Jan P.M.
AU - Cui, Li
AU - Zou, Wen Quan
N1 - Funding Information:
Funding: This research was funded in part by National Institutes of Health (R01NS109532) to W.-Q.Z., CJD Foundation to W.-Q.Z., Centers for Disease Control and Prevention grant underneath funding, and ALZ/ARUK/MJFF/Weston to W.-Q.Z, as well as the National Natural Science Foundation of China (NNSFC) (No. 81671186) to L.C.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/5
Y1 - 2021/5
N2 - Prion is an infectious protein (PrPSc ) that is derived from a cellular glycoprotein (PrPC ) through a conformational transition and associated with a group of prion diseases in animals and humans. Characterization of proteinase K (PK)-resistant PrPSc by western blotting has been critical to diagnosis and understanding of prion diseases including Creutzfeldt-Jakob disease (CJD) and Gerstmann-Sträussler-Scheinker (GSS) disease in humans. However, formation as well as biochemical and biological properties of the glycoform-selective PrPSc in variably protease-sensitive prionopathy (VPSPr) remain poorly understood. Here we reveal that formation of the ladder-like PrPSc in VPSPr is a PK-dependent two-step process, which is enhanced by basic pH. Two sets of PrPSc fragments can be identified with antibodies directed against an intermediate or a C-terminal domain of the protein. Moreover, antibodies directed against specific PrP glycoforms reveal faster electrophoretic migrations of PrP fragments mono-glycosylated at residue 181 and 197 in VPSPr than those in sporadic CJD (sCJD). Finally, RT-QuIC assay indicates that PrPSc-seeding activity is lower and its lag time is longer in VPSPr than in sCJD. Our results suggest that the glycoform-selective PrPSc in VPSPr is associated with altered glycosylation, resulting in different PK-truncation and aggregation seeding activity compared to PrPSc in sCJD.
AB - Prion is an infectious protein (PrPSc ) that is derived from a cellular glycoprotein (PrPC ) through a conformational transition and associated with a group of prion diseases in animals and humans. Characterization of proteinase K (PK)-resistant PrPSc by western blotting has been critical to diagnosis and understanding of prion diseases including Creutzfeldt-Jakob disease (CJD) and Gerstmann-Sträussler-Scheinker (GSS) disease in humans. However, formation as well as biochemical and biological properties of the glycoform-selective PrPSc in variably protease-sensitive prionopathy (VPSPr) remain poorly understood. Here we reveal that formation of the ladder-like PrPSc in VPSPr is a PK-dependent two-step process, which is enhanced by basic pH. Two sets of PrPSc fragments can be identified with antibodies directed against an intermediate or a C-terminal domain of the protein. Moreover, antibodies directed against specific PrP glycoforms reveal faster electrophoretic migrations of PrP fragments mono-glycosylated at residue 181 and 197 in VPSPr than those in sporadic CJD (sCJD). Finally, RT-QuIC assay indicates that PrPSc-seeding activity is lower and its lag time is longer in VPSPr than in sCJD. Our results suggest that the glycoform-selective PrPSc in VPSPr is associated with altered glycosylation, resulting in different PK-truncation and aggregation seeding activity compared to PrPSc in sCJD.
KW - Creutzfeldt-Jakob disease (CJD)
KW - Gerstmann-Sträussler-Scheinker (GSS)
KW - Glycoform-selective prion formation
KW - Prions disease
KW - Real-time quaking-induced conversion (RT-QuIC) assay
KW - Variably protease-sensitive prionopathy (VPSPr)
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U2 - 10.3390/pathogens10050513
DO - 10.3390/pathogens10050513
M3 - Article
AN - SCOPUS:85105295080
SN - 2076-0817
VL - 10
JO - Pathogens
JF - Pathogens
IS - 5
M1 - 513
ER -
