title: Insulin-stimulating peptide from a tryptic digest of bovine serum albumin: purification and characterization.
abstract: A procedure was established for isolation of a low molecular weight polypeptide with insulin-stimulating activity in apparent homogeneity from a tryptic digest of bovine serum albumin on a semipreparative scale. Purification of this insulin-stimulating peptide (ISP) was monitored by an adipose-explant assay in which stimulation of fatty acid synthesis from glucose by insulin was measured. The polypeptide was purified by a combination of DEAE-cellulose column chromatography, gel filtration on Bio-Gel P-10, hydrophobic chromatography on a semipreparative C18 reversed-phase HPLC column, and ion exchange chromatography on an SP-5PW HPLC column. The primary structure of ISP was deduced. ISP is a two-chain polypeptide consisting of 71 amino acid residues, and corresponds essentially to residues 115-143 and 144-184 (185) of bovine serum albumin connected to each other by a disulfide bridge. But comparison of the sequence of ISP with that of the relevant regions of bovine serum albumin determined by Brown indicated the presence of one tyrosine insertion between residues 155 and 156 of albumin. Therefore, the molecular weight of ISP was calculated to be 8,496.
mesh_terms: D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D005227:Fatty Acids / Q000096:biosynthesis; D005947:Glucose / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D009924:Organ Culture Techniques; D010455:Peptides / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine; D014357:Trypsin
chemical_list: D000596:Amino Acids; D005227:Fatty Acids; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C047279:insulin-stimulating peptide, human; D012710:Serum Albumin, Bovine; D014357:Trypsin; D005947:Glucose