Transfer of overexpressed protein into organelle and its regulation mechanism

点击翻译按钮获取中文摘要

cDNA encoding mitochondrial very-long-chain acyl-CoA dehydrogenase was cloned. Amino acid sequence indicated that the enzyme had about 180 amino acids of extra polypeptide at the carboxyl terminal side, when it was compared with the other acyl-CoA dehydrogenases. A series of mutants, deleting about 60 amino acids of polypeptide from carboxyl terminus, were inserted into vaccinia virus to construct recombinant viruses. Expressed protein in rat hepatoma H4 was examined by means of immunoblot analysis. Mutant protein was less stable in mitochondria than mature protein concerning period after the expression. Mutant protein also lost the features of membrane binding and association to form dimer. These results indicate that the extra polypeptide at carboxyl side has unique function to stabilize the protein in mitochondria. Similar results are obtained concerning the extra polypeptide at carboxyl side of peroxisomal acyl-CoA oxidase.