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.
| Language | English |
|---|---|
| Pages | 5456-5469 |
| Journal | Molecular Neurobiology |
| Volume | 56 |
| Issue number | 8 |
| Early online date | 5 Jan 2019 |
| DOIs | |
| Publication status | Published - Aug 2019 |
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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
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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 journal › Article › Academic › peer-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
IS - 8
ER -