Proteomic analysis of liver plasma membrane from hepatitis B surface antigen transgenic mice
10.3760/cma.j.issn.1000-6680.2012.10.001
- VernacularTitle:转乙型肝炎病毒表面抗原基因小鼠肝细胞质膜蛋白质组学研究
- Author:
Xiaofang JIA
;
Chunhong LI
;
Xia PENG
;
Lin YIN
;
Yanling FENG
;
Fang MA
;
Lijun ZHANG
- Publication Type:Journal Article
- Keywords:
Mouse,transgenic;
Hepatitis B surface antigen;
Viral envelope proteins;
Proteomics
- From:
Chinese Journal of Infectious Diseases
2012;30(10):577-582
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the differential liver plasma membrane (PM) proteins that may be related to the occurrence,development and reversal process of hepatitis and to understand the pathogenesis of hepatitis and the new drug targets by performing a comparative proteomics research of liver PM between hepatitis B surface antigen (HBsAg) transgenic mice and wild-type C57 mice.Methods A 6-month-old HBsAg transgenic mouse model was established.The pathological examination was performed to observe the pathological changes of transgenic mice and wild-type C57 mice.The PM from liver tissue of 6-month-old transgenic mouse and the control mouse were purified through twice sucrose density grade centrifugation combined with second antibody magnetic bead enrichment.The purity of extracted PM was verified by Western blot.Differential proteome expression analysis was performed by using two-dimensional electrophoresis (2-DE) and ImageMaster software analysis.The differentially expressed proteins were lysed by trypsin and identified by liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) analysis.Results The pathological examination results showed that hepatitis was observed in the transgenic mouse group,while no abnormity was found in the controls.The PM was successfully enriched and the mitochondria contamination was reduced by sucrose density grade centrifugation combined with second antibody magnetic bead purification treatment.Thirty differential mice liver PM protein spots were visualized,in which 11 non-redundant proteins were successfully identified by LC-MS/MS in transgenic mouse group,including 9 up regulated protein spots and 2 down-regulated protein spots.These differentially expressed proteins included keratin,cardiac Ca2+ release channel,cytochrome B5,ATP synthase subunit alpha,etc.Conclusions A batch of HBsAg gene expression related differential proteins are identified in mouse liver plasma.These proteins might be new drug targets for anti-HBV treatment.This study will guide further investigation on the mechanism of HBV infection induced hepatitis.
