In Vitro Seeding Activity of Glycoform-Deficient Prions from Variably Protease-Sensitive Prionopathy and Familial CJD Associated with PrPV180I Mutation

Zerui Wang, Jue Yuan, Pingping Shen, Romany Abskharon, Yue Lang, Johnny Dang, Alise Adornato, Ling Xu, Jiafeng Chen, Jiachun Feng, Mohammed Moudjou, Tetsuyuki Kitamoto, Jan Langeveld, Brian Appleby, Jiyan Ma, Qingzhong Kong, Robert B. Petersen, Wen Quan Zou, Li Cui

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Both sporadic variably protease-sensitive prionopathy (VPSPr) and familial Creutzfeldt-Jakob disease linked to the prion protein (PrP) V180I mutation (fCJDV180I) have been found to share a unique pathological prion protein (PrPSc) that lacks the protease-resistant PrPSc glycosylated at residue 181 because two of four PrP glycoforms are apparently not converted into the PrPSc from their cellular PrP (PrPC). To investigate the seeding activity of these unique PrPSc molecules, we conducted in vitro prion conversion experiments using serial protein misfolding cyclic amplification (sPMCA) and real-time quaking-induced conversion (RT-QuIC) assays with different PrPC substrates. We observed that the seeding of PrPSc from VPSPr or fCJDV180I in the sPMCA reaction containing normal human or humanized transgenic (Tg) mouse brain homogenates generated PrPSc molecules that unexpectedly exhibited a dominant diglycosylated PrP isoform along with PrP monoglycosylated at residue 181. The efficiency of PrPSc amplification was significantly higher in non-CJDMM than in non-CJDVV human brain homogenate, whereas it was higher in normal TgVV than in TgMM mouse brain homogenate. PrPC from the mixture of normal TgMM and Tg mouse brain expressing PrPV180I mutation (Tg180) but not TgV180I alone was converted into PrPSc by seeding with the VPSPr or fCJDV180I. The RT-QuIC seeding activity of PrPSc from VPSPr and fCJDV180I was significantly lower than that of sCJD. Our results suggest that the formation of glycoform-selective prions may be associated with an unidentified factor in the affected brain and the glycoform-deficiency of PrPSc does not affect the glycoforms of in vitro newly amplified PrPSc.

LanguageEnglish
Pages5456-5469
JournalMolecular Neurobiology
Volume56
Issue number8
Early online date5 Jan 2019
DOIs
Publication statusPublished - Aug 2019

Fingerprint

Prions
Peptide Hydrolases
Mutation
Brain
Transgenic Mice
Creutzfeldt-Jakob Syndrome
In Vitro Techniques
Prion Proteins
Protein Isoforms
Proteins

Keywords

  • Humanized transgenic mice
  • Polymorphism
  • Prion
  • Prion disease
  • Real-time quaking-induced conversion (RT-QuIC)
  • Serial protein misfolding cyclic amplification (sPMCA)
  • Variably protease-sensitive prionopathy (VPSPr)

Cite this

Wang, Zerui ; Yuan, Jue ; Shen, Pingping ; Abskharon, Romany ; Lang, Yue ; Dang, Johnny ; Adornato, Alise ; Xu, Ling ; Chen, Jiafeng ; Feng, Jiachun ; Moudjou, Mohammed ; Kitamoto, Tetsuyuki ; Langeveld, Jan ; Appleby, Brian ; Ma, Jiyan ; Kong, Qingzhong ; Petersen, Robert B. ; Zou, Wen Quan ; Cui, Li. / In Vitro Seeding Activity of Glycoform-Deficient Prions from Variably Protease-Sensitive Prionopathy and Familial CJD Associated with PrPV180I Mutation. In: Molecular Neurobiology. 2019 ; Vol. 56, No. 8. pp. 5456-5469.
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abstract = "Both sporadic variably protease-sensitive prionopathy (VPSPr) and familial Creutzfeldt-Jakob disease linked to the prion protein (PrP) V180I mutation (fCJDV180I) have been found to share a unique pathological prion protein (PrPSc) that lacks the protease-resistant PrPSc glycosylated at residue 181 because two of four PrP glycoforms are apparently not converted into the PrPSc from their cellular PrP (PrPC). To investigate the seeding activity of these unique PrPSc molecules, we conducted in vitro prion conversion experiments using serial protein misfolding cyclic amplification (sPMCA) and real-time quaking-induced conversion (RT-QuIC) assays with different PrPC substrates. We observed that the seeding of PrPSc from VPSPr or fCJDV180I in the sPMCA reaction containing normal human or humanized transgenic (Tg) mouse brain homogenates generated PrPSc molecules that unexpectedly exhibited a dominant diglycosylated PrP isoform along with PrP monoglycosylated at residue 181. The efficiency of PrPSc amplification was significantly higher in non-CJDMM than in non-CJDVV human brain homogenate, whereas it was higher in normal TgVV than in TgMM mouse brain homogenate. PrPC from the mixture of normal TgMM and Tg mouse brain expressing PrPV180I mutation (Tg180) but not TgV180I alone was converted into PrPSc by seeding with the VPSPr or fCJDV180I. The RT-QuIC seeding activity of PrPSc from VPSPr and fCJDV180I was significantly lower than that of sCJD. Our results suggest that the formation of glycoform-selective prions may be associated with an unidentified factor in the affected brain and the glycoform-deficiency of PrPSc does not affect the glycoforms of in vitro newly amplified PrPSc.",
keywords = "Humanized transgenic mice, Polymorphism, Prion, Prion disease, Real-time quaking-induced conversion (RT-QuIC), Serial protein misfolding cyclic amplification (sPMCA), Variably protease-sensitive prionopathy (VPSPr)",
author = "Zerui Wang and Jue Yuan and Pingping Shen and Romany Abskharon and Yue Lang and Johnny Dang and Alise Adornato and Ling Xu and Jiafeng Chen and Jiachun Feng and Mohammed Moudjou and Tetsuyuki Kitamoto and Jan Langeveld and Brian Appleby and Jiyan Ma and Qingzhong Kong and Petersen, {Robert B.} and Zou, {Wen Quan} and Li Cui",
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Wang, Z, Yuan, J, Shen, P, Abskharon, R, Lang, Y, Dang, J, Adornato, A, Xu, L, Chen, J, Feng, J, Moudjou, M, Kitamoto, T, Langeveld, J, Appleby, B, Ma, J, Kong, Q, Petersen, RB, Zou, WQ & Cui, L 2019, 'In Vitro Seeding Activity of Glycoform-Deficient Prions from Variably Protease-Sensitive Prionopathy and Familial CJD Associated with PrPV180I Mutation', Molecular Neurobiology, vol. 56, no. 8, pp. 5456-5469. https://doi.org/10.1007/s12035-018-1459-0

In Vitro Seeding Activity of Glycoform-Deficient Prions from Variably Protease-Sensitive Prionopathy and Familial CJD Associated with PrPV180I Mutation. / Wang, Zerui; Yuan, Jue; Shen, Pingping; Abskharon, Romany; Lang, Yue; Dang, Johnny; Adornato, Alise; Xu, Ling; Chen, Jiafeng; Feng, Jiachun; Moudjou, Mohammed; Kitamoto, Tetsuyuki; Langeveld, Jan; Appleby, Brian; Ma, Jiyan; Kong, Qingzhong; Petersen, Robert B.; Zou, Wen Quan; Cui, Li.

In: Molecular Neurobiology, Vol. 56, No. 8, 08.2019, p. 5456-5469.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - In Vitro Seeding Activity of Glycoform-Deficient Prions from Variably Protease-Sensitive Prionopathy and Familial CJD Associated with PrPV180I Mutation

AU - Wang, Zerui

AU - Yuan, Jue

AU - Shen, Pingping

AU - Abskharon, Romany

AU - Lang, Yue

AU - Dang, Johnny

AU - Adornato, Alise

AU - Xu, Ling

AU - Chen, Jiafeng

AU - Feng, Jiachun

AU - Moudjou, Mohammed

AU - Kitamoto, Tetsuyuki

AU - Langeveld, Jan

AU - Appleby, Brian

AU - Ma, Jiyan

AU - Kong, Qingzhong

AU - Petersen, Robert B.

AU - Zou, Wen Quan

AU - Cui, Li

PY - 2019/8

Y1 - 2019/8

N2 - Both sporadic variably protease-sensitive prionopathy (VPSPr) and familial Creutzfeldt-Jakob disease linked to the prion protein (PrP) V180I mutation (fCJDV180I) have been found to share a unique pathological prion protein (PrPSc) that lacks the protease-resistant PrPSc glycosylated at residue 181 because two of four PrP glycoforms are apparently not converted into the PrPSc from their cellular PrP (PrPC). To investigate the seeding activity of these unique PrPSc molecules, we conducted in vitro prion conversion experiments using serial protein misfolding cyclic amplification (sPMCA) and real-time quaking-induced conversion (RT-QuIC) assays with different PrPC substrates. We observed that the seeding of PrPSc from VPSPr or fCJDV180I in the sPMCA reaction containing normal human or humanized transgenic (Tg) mouse brain homogenates generated PrPSc molecules that unexpectedly exhibited a dominant diglycosylated PrP isoform along with PrP monoglycosylated at residue 181. The efficiency of PrPSc amplification was significantly higher in non-CJDMM than in non-CJDVV human brain homogenate, whereas it was higher in normal TgVV than in TgMM mouse brain homogenate. PrPC from the mixture of normal TgMM and Tg mouse brain expressing PrPV180I mutation (Tg180) but not TgV180I alone was converted into PrPSc by seeding with the VPSPr or fCJDV180I. The RT-QuIC seeding activity of PrPSc from VPSPr and fCJDV180I was significantly lower than that of sCJD. Our results suggest that the formation of glycoform-selective prions may be associated with an unidentified factor in the affected brain and the glycoform-deficiency of PrPSc does not affect the glycoforms of in vitro newly amplified PrPSc.

AB - Both sporadic variably protease-sensitive prionopathy (VPSPr) and familial Creutzfeldt-Jakob disease linked to the prion protein (PrP) V180I mutation (fCJDV180I) have been found to share a unique pathological prion protein (PrPSc) that lacks the protease-resistant PrPSc glycosylated at residue 181 because two of four PrP glycoforms are apparently not converted into the PrPSc from their cellular PrP (PrPC). To investigate the seeding activity of these unique PrPSc molecules, we conducted in vitro prion conversion experiments using serial protein misfolding cyclic amplification (sPMCA) and real-time quaking-induced conversion (RT-QuIC) assays with different PrPC substrates. We observed that the seeding of PrPSc from VPSPr or fCJDV180I in the sPMCA reaction containing normal human or humanized transgenic (Tg) mouse brain homogenates generated PrPSc molecules that unexpectedly exhibited a dominant diglycosylated PrP isoform along with PrP monoglycosylated at residue 181. The efficiency of PrPSc amplification was significantly higher in non-CJDMM than in non-CJDVV human brain homogenate, whereas it was higher in normal TgVV than in TgMM mouse brain homogenate. PrPC from the mixture of normal TgMM and Tg mouse brain expressing PrPV180I mutation (Tg180) but not TgV180I alone was converted into PrPSc by seeding with the VPSPr or fCJDV180I. The RT-QuIC seeding activity of PrPSc from VPSPr and fCJDV180I was significantly lower than that of sCJD. Our results suggest that the formation of glycoform-selective prions may be associated with an unidentified factor in the affected brain and the glycoform-deficiency of PrPSc does not affect the glycoforms of in vitro newly amplified PrPSc.

KW - Humanized transgenic mice

KW - Polymorphism

KW - Prion

KW - Prion disease

KW - Real-time quaking-induced conversion (RT-QuIC)

KW - Serial protein misfolding cyclic amplification (sPMCA)

KW - Variably protease-sensitive prionopathy (VPSPr)

U2 - 10.1007/s12035-018-1459-0

DO - 10.1007/s12035-018-1459-0

M3 - Article

VL - 56

SP - 5456

EP - 5469

JO - Molecular Neurobiology

T2 - Molecular Neurobiology

JF - Molecular Neurobiology

SN - 0893-7648

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ER -