Proteasomal inhibitor induces PINK1 aggresome formation and aggregating features
- VernacularTitle:蛋白酶体抑制剂诱导帕金森病PINK1蛋白聚集物的形成与特征
- Author:
Yu-Hu ZHANG
;
Bei-Sha TANG
;
Lu WEN
;
Bo XU
;
Jian-Guang TANG
;
Ji-Feng GUO
;
Kun XIA
;
Lu SHEN
;
Hong JIANG
;
- Publication Type:Journal Article
- Keywords:
Parkinson disease;
Protein kinases;
Leupeptins;
Inclusion bodies
- From:
Chinese Journal of Neurology
2000;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the PINK1 aggresome formation and it's features in response to proteasomal inhibition.Methods Full-length PINK1 cDNA were amplified by polymerase chain reaction (PCR)from fetus brain cDNA library and subcloned into the EcoR I and BamH I sites of the vector pEGFP- N1.The integrity of the constructs was confirmed by sequencing.COS-7 cells were transiently transfected with PINK1-pEGFP-N1 using Lipofectamine 2000.Cells were treated by MG-132 in order to test the effect of proteasome inhibition on aggregation formation.The protein level of wild-type PINK1 with or without MG-132 treatment was confirmed by Western blot analysis.The formation of PINK1 aggregates was tested by fluorescence and the presence of ubiquitin,and ?-synuclein in PINK1 aggregates was examined by immunofluorescence and confocal microscopy.Results The expression level of PINK1 was significant increased into the form of aggregate in cells treated with MG-132;immunostaining for endogenous ubiquitin and ?-synuclein revealed a co-localization of both proteins in PINK1-positive aggregates.Conclusions In the presence of MG-132,overexpressed PINK1 forms into aggregates,whose components are ubiquitin and ?-synuclein.
