A Preliminary Study On The Function Of L-Caveolin-like Protein

Caveolae is the surface of the cell membrane specific depression,mainly composed of lipids and proteins,lipids including cholesterol,glycolipids,phospholipids;protein is mainly the Caveolin family.Caveolin-1 protein in mammals is involved in cell metabolism,signal transduction and tumor cell bi-directional regulation.In this paper,the full-length sequence of L-Caveolin-like gene was obtained by 5 'and 3'Race rapid amplification technique.The full length of the gene was 1817 bp,the 5'-UTR was 253 bp,the 3'-UTR was 784 bp,the ORF was 780 bp and codes 259 amino acids.The structural characteristics,physical and chemical properties of L-Caveolin-like were predicted by bioinformatics-related software.According to the homology of other species Caveolin protein and L-Caveolin-like protein,phylogenetic tree was made to determine its position in the phylogenetic development of each species.L-Caveolin-like was cloned to p ET-30 a prokaryotic expression vector and transformed into E.coli to induce,express and purify L-Caveolin-like protein,then New Zealand rabbits were immuned with L-Caveolin-like protein and antibody was purified by Protein-G affinity chromatography from antiserum.A high specificity of the lampreys L-Caveolin-like rabbit polyclonal antibody was prepared.Immunohistochemistry assay showed that L-Caveolin-like was expressed in the intestine,liver,kidney,gill,sp-body,heart.Importantly the expression level of L-Caveolin-like in intestinal and liver tissues was relatively high.The eukaryotic expression vector of p EGFP-N1-L-Caveolin-like was constructed and transfected into Hela cells by liposomes.Ultra-high resolution microscopy showed that L-Caveolin-like was located in mitochondria of Hela cells.The expression of CO?,Cyt C and PUMA gene was decreased by real-time quantitative PCR,L-Caveolin-like was used to study the effects of L-Caveolin-like on mitochondrial oxidative stress and anti-apoptosis;Increased cellular ATP levels suggest that it has the effect of maintaining mitochondrial production to prevent mitochondrial damage;The expression of L-Caveolin-like in response to LPS-stimulated lamprey was significantly higher,suggesting that L-Caveolin-like could protect the lamprey infected with exogenous pathogens;In addition,the overexpression of L-Caveolin-like can up-regulate the expression of EGF,FGF and PDGF,suggesting that L-Caveolin-like can involve in proliferation.In summary,L-Caveolin-like plays an important role in promoting cell proliferation,preventing mitochondrial oxidative stress and inhibiting apoptosis,but its specific mechanism remains to be further studied.