Recombinant expression of an insulin-like peptide 3 (INSL3) precursor and its enzymatic conversion to mature human INSL3

摘要: Insulin-like peptide 3 (INSL3), which is primarily expressed in the Leydig cells of testes, a member insulin superfamily hormones. One its primary functions to initiate and mediate descent testes male fetus via interaction with G protein-coupled receptor, RXFP2. Study has relied upon chemical synthesis separate A- B-chains subsequent chain recombination. To establish an alternative approach preparation human INSL3, we designed recombinantly single-chain INSL3 precursor Escherichia coli cells. The was solubilized from inclusion body, purified almost homogeneity by immobilized metal-ion affinity chromatography refolded efficiently in vitro. subsequently converted mature sequential endoproteinase Lys-C carboxypeptidase B treatment. CD spectroscopic analysis mapping showed that possessed insulin-like fold expected disulfide linkages. Recombinant demonstrated full activity stimulating cAMP RXFP2-expressing Interestingly, comparable two-chain suggesting receptor-binding region within mid- C-terminal B-chain maintained active conformation precursor. This study not only provides efficient for preparation, but also resulted acquisition useful template additional structural functional studies peptide.