Proteomic Characterization of Human Erythrocyte 20S Proteasome and Analysis of Species-dependent 20S Proteasome Heterogeneity
- VernacularTitle:人红细胞20S蛋白酶体的蛋白质组学表征及不同来源20S蛋白酶体异质性的研究
- Author:
Guoqiang CHEN
;
Hui LIU
;
Haijing ZHANG
;
Yanchun DENG
;
Zhili LI
- Publication Type:Journal Article
- Keywords:
Proteasome;
heterogeneity;
nondenaturing polyacrylamide gel electrophoresis;
differential centrifugation;
cell line
- From:
Chinese Journal of Analytical Chemistry
2009;37(12):1711-1716
- CountryChina
- Language:Chinese
-
Abstract:
A method was developed for purification of 20S proteasome (20S core particle, CP) by combining differential centrifugations with nondenaturing polyacrylamide gel electrophoresis (native-PAGE), irrespective of species origins of CPs. CP purified from human erythrocytes was subjected to proteomic analysis by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), revealing 33 spots of subunit isoforms with different molecular weights and isoelectric points, more than 14 constituent subunits. Furthermore, other four CPs were purified from yeast, mouse liver, two pancreatic cancer cell lines SW1990 and PANC-1 using this method mentioned above, and subjected to proteasome heterogeneity analysis by native/SDS-PAGE (native/sodium dodecyl sulphate polyacrylamide gel electrophoresis), together with CP from erythrocytes. The method described acts as a rapid and effective tool for CP isolations, and the results obtained may be served as a footstone for the investigations of species-dependent proteasome heterogeneity.
