Study On Candidate Interacting Proteins With TCTP Protein

Objective:To screen the candidate proteins of hepatocyte which interact with TCTP protein.Methods: TCTPwas amplified by polymerase chain reaction (PCR) from cDNA of hepatoma carcinoma cell. Bait plasmid expressing TCTP was re-constructed by cloning the target region into pDEST32 and named pDEST32-TCTP. Cotransform pDEST32- TCTP and prey plasmids which expressing liver cDNA library into the yeast cell MaV203 by Liac-mediated transformation. Diploid yeast was plated on synthetic dropout nutrient medium for selection. Prey plasmids were extracted from positive colonies and the inserted cDNA were sequenced for bioinformatics analysis. We applied the GST and Co-IP method to validate the interaction between the TCTP protein and candidate proteins.Results:By yeast two-hybrid method, six colonies were screened out as positive colonies and their inserted cDNA were sequenced. Sequences of positive colonies code Smad4,HSP60,TAKl,Bcl-xl,FS,Skp2. GST and Co-IP proved TCTP protein can interact with Skp2 and FS proteins.Conclusion:According to the yeast two-hybrid results, six polypeptides/proteins were proved to be candidate interacting proteins with TCTP protein. This research indicates that TCTP protein may involve in tumor genesis and development. However, the roles of TCTP protein may be comprehensive and remain to be elucidated. Nevertheless, our work offers a new detail to probe the role of TCTP protein in the pathogenesis of liver fibrosis and hepatic celluar carcinoma.