ABSTRACT
A região PreS1 da proteína L é importante na adesão celular e, consequentemente, o vírus da hepatite B (HBV) infectividade. Para identificar novas proteínas que interagem PreS1 , realizamos Glutationa -S -transferase (GST) suspenso, electroforese bidimensional do gel ( 2-DE) e os testes de espectrometria de massa. proteínas Glucose- regulamentado (GRP ) 78 e 75 foram encontrados para vincular PreS1 . As interações entre PreS1 e GRP75 foram confirmados por uma experiência de co - imunoprecipitação . GRP78 e GRP75 pode desempenhar um papel importante na mediação HBV virion entrar em hepatócitos e regulação dobradura apropriada da proteína L , devido às suas funções críticas de enovelamento de proteínas e tráfico. A descoberta do romance PreS1 proteína enriquece nosso conhecimento sobre o mecanismo molecular da infecção pelo HBV.
The PreS1 region of the L protein is important in cell attachment and consequently in hepatitis B virus (HBV) infectivity. To identify novel PreS1 interacting protein, we performed Glutathione-S-transferase (GST) pull-down, two-dimensional gel electrophoresis (2-DE) and mass spectrometry assays. Glucose-regulated proteins (GRP) 78 and 75 were found to bind PreS1. The interactions between PreS1 and GRP75 were confirmed by a co-immunoprecipitation experiment. GRP78 and GRP75 may play important roles in mediating HBV virion entering into hepatocyte and regulating proper folding of the L protein due to their critical functions in protein folding and trafficking. The finding of novel PreS1 binding protein enriches our knowledge about molecular mechanism of HBV infection.
Subject(s)
Humans , Glutathione S-Transferase pi , Hepatocytes , In Vitro Techniques , Mass Spectrometry , Polymerase Chain Reaction , Proteomics , Hepatitis B virus/isolation & purification , Electrophoresis , Methods , MethodsABSTRACT
Objective To identify the effect of glucose regulating protein 75 expression in PC12 cells under glucose deprivation.Methods MTT method was used to monitor the cell viability.The leakage of LDH was represented as the index of cell injury.Flow cytometry was applied for cell cycle analysis,apoptosis and necrosis.Using RT-PCR detected and quantified mRNA.Western blot to measure relative amounts of the protein.Results Cell viability decreased and the LDH leakage increased.While the treated PC12 cells had two main forms of apoptosis and necrosis.GRP75 mRNA and protein levels were upregulated.Conclusion GRP75 has protective function during glucose deprivation.