Isolation and a partial amino acid sequence of insulin from the islet tissue of cod (Gadus callarias)

Abstract

Insulin was isolated by gel filtration and ion-exchange chromatography from extracts of the discrete islet tissue of cod. The final preparation yielded a single band on electrophoresis at 2 pH values. The biological potency was 11.5 international units/mg in mouse-convulsion and other assay procedures. Glycine and methionine are N-terminal amino acids of the A and B chains, respectively. An estimate of the molecular weight together with amino acid analyses indicated that cod insulin, like the bovine hormone, consists of 51 amino acid residues. In contrast, the amino acid composition differs markedly from bovine insulin. Oxidation of insulin with performic acid yielded the A and B peptide chains, which were separated by ion-exchange chromatography. Sequence studies on smaller peptides isolated from enzymic digests or from dilute acetic acid hydrolysates of the 2 chains have established the sequential order of 14 of the 21 amino acid residues of the A chain and 25 of the 30 amino acid residues of the B chain.