Comprehensive proteomic analysis of host cell lipid rafts modified by HBV infection |
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Authors: | Xie Na Huang Kai Zhang Tao Lei Yunlong Liu Rui Wang Kui Zhou Shengtao Li Jingyi Wu Jinhua Wu Hong Deng Cao Zhao Xia Nice Edouard Collins Huang Canhua |
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Affiliation: | a The State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610041, PR Chinab The School of Biomedical Sciences, Chengdu Medical College, Chengdu, 610083, PR Chinac Department of Hepatobiliary Pancreatic Surgery, West China Hospital, Sichuan University, Chengdu, 610041, PR Chinad Faculty of Medecine, Nursing and Health Sciences, Monash University, Clayton, Victoria 3800, Australia |
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Abstract: | Lipid rafts are cholesterol- and sphingolipid-rich membrane microdomains that have been shown to participate in the entry, assembly and budding of various viruses. However, their involvement in HBV replication remains poorly characterized. In a preliminary study, we observed that HBV release could be markedly impaired by methyl-β-cyclodextrin mediated depletion of cholesterol in lipid rafts, and that this effect could be reversed by replenishment of exogenous cholesterol, suggesting that lipid rafts play an important role in the HBV life cycle. To further understanding how HBV exploited host cell lipid rafts to benefit replication, comprehensive proteomic approaches were used to profile the proteome changes of host cell lipid rafts in response to HBV infection using 2DE-MS/MS, in combination with SILAC-based quantitative proteomics. Using these approaches, a total of 97 differentially expressed proteins were identified. Bioinformatics analysis suggested that multiple host cell pathways were involved in the HBV infection processes including signal transduction, metabolism, immune response, transport, vesicle trafficking, cell adhesion and cellular ion homeostasis. These data will provide valuable clues for further investigation of HBV pathogenesis. |
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Keywords: | 2DE, 2 dimensional polyacrylamide gel electrophoresis CAV1, caveolin 1 DMEM, Dulbecco's modified eagle medium DMSO, dimethyl sulfoxide DRMs, detergent-resistant membranes ER, endoplasmic reticulum ESI-Q-TOF, electrospray ionization quadrupole time-of-flight GOEAST, Gene Ontology Enrichment Analysis Software Toolkit HBsAg, hepatitis B surface antigen HBV, hepatitis B virus HCC, hepatocellular carcinoma HIV, human immunodeficiency virus HSV, herpes simplex virus IPA, Ingenuity Pathways Analysis MLV, murine leukemia virus MS, mass spectrometry MTT, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide MβCD, methyl-β-cyclodextrin NDV, Newcastle disease virus PID, Pathway Interaction Database RSV, respiratory syncytial virus SILAC, stable isotope labeling with amino acids in cell culture SREBP1c, nuclear sterol regulatory element-binding protein 1c SV40, simian virus 40 SVPs, subviral particles TEM, transmission electron microscope TfR1, transferrin receptor 1 |
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