Separation and identification of hsp27 interacting proteins from human hepatocellular carcinoma cells / 重庆医科大学学报

ABSTRACT

Objective:To find out heatshock protein HSP27 interactingproteins in the hepatocellularcarcinoma(HCC).Methods:HSP27 interacting proteins in the human HCC HepG2.2215 cells were seperated and identified.HSP27 binding proteins were recovered by antiHSP27 antibody immunoprecipitation with the total proteins from HCC HepG2.2215 cells.After in-solution digested and analyzed by LC-ESI-ITMS/MS,the recovered protein complexes were identified by peptide sequence tags(PST)and database searching,and were confimed by immunoprecipitation and Western blot analysis.Results:Seven HSP27 binding proteins were firstly identified by database searching,which were HSP90,GRP-78 and HSP70 of HSP70 family members,lysyl hydroxylase,Keratin 9,Keratin 10,Keratin 1.The identified seven HSP27-interacting proteins by IP coupled with MS could be classified into three functional categories based on their functionschaperones,cytoskeleton organization,metabolic enzymes.Conclusion:HSP27 in HCC is a related proteins involved in the regulation,by interacting with some peptides,or small molecule protein to control the tumor development.