The effect of proteasomal inhibitors on anterior pharynx decfective-1 expression in neuronal cells
10.3760/cma.j.issn.1006-7876.2010.11.016
- VernacularTitle:蛋白酶体抑制剂对神经细胞前咽缺陷蛋白-1表达的影响
- Author:
Kejian WANG
;
Zhimin LONG
;
Baobing GAO
;
Lei ZHAO
;
Weitian LU
;
Shengwei GAN
;
Guiqiong HE
- Publication Type:Journal Article
- Keywords:
Neurons;
Membrane proteins;
Cysteine proteinase inhibitors
- From:
Chinese Journal of Neurology
2010;43(11):795-800
- CountryChina
- Language:Chinese
-
Abstract:
Objectives To investigate whether degradation of anterior pharynx decfective-1(Aph-1) goes through proteasomal pathway or lysosomal pathway.Methods Various methods such as cell culture,Western blotting,pulse-chase metabolic labeling technique,double immunofluoresecnt staining,combined with proteasomal and lysosomal inhibition were used to check Aph-1 expression level in stable Aph-1-transfected or non-transfected neuronal(SH-SY5Y)cell line.Results Using Western blotting,treating the neuronal cells with proteasome specific inhibitors significantly increased the expression of both endogenous and exogenous Aph-1.The effect of the proteasome inhibitors on Aph-1 expression was dose-and time-dependent Lysosomal pathway was not involved in Aph-1 degradation. Pulse-chase metabolic labeling experiment showed that the turnover of newly-synthesized radiolabeled Aph-1 protein was blocked by Lactacystin.Double immunofluorescent staining revealed colocalization of Aph-1 and ubiquitin in the same cells.Conclusion Degradation of Aph-1 protein is mediated by proteasomal pathway in neuronal cells,and is not related to lysosomal pathway.Aph-1 protein is ubiquitinated before degradation.
