Interaction of SARP1 proteins in fibroblasts of hypertrophic scar / 中华医学美学美容杂志

ABSTRACT

Objective To study interaction proteins with secreted apoptosis-related protein 1 (SARP1) in fibroblasts of hypertrophic scar and to analyze its molecular mechanisms.Methods The recombinant adenovirus Ad-SARP1 was successfully constructed and transfected into the fibroblasts of hypertrophic scar in culture.The proteins were precipitated by immunoprecipitation and separated by SDS-PAGE,then it was stained with Coomassie blue and proteins from SDS-PAGE gel electrophoresis strip were analyzed with enzymolysis and mass spectrometric in turn.The peptide sequences were obtained according to mass spectrometry and the database were searched automatically.Results The results showed that in control cells,Ad-SARP1 and Ad-EGFP infected cells,were precipitated 7 protein bands,their molecular weights were about 93 × 103,43 × 103,40 × 103,37 × 103,31 × 103,26 × 103 and 12× 103,respectively; without SARP1 antibody the protein bands did not precipitate.Analysis of the 6 protein bands showed that proteins that might interact with SARP1 included periostin precursor (OSF-2),asporin precursor (PLAP1),phosphoglycerate kinase 1,rCG50690,apolipoprotein A-I precursor (Apo-AI),and thioredoxin 1 (TRX1).Conclusions The interaction proteins of SARP1 can be obtained by immunoprecipitation combined with liquid chromatography/mass spectrometry and ion trap detection technology.These results provide new clues for the mechanism of SARP1 regulates the apoptosis signal pathway of HSFb.