title: Insulin-binding peptide. Design and characterization.
abstract: The design and characterization of a six-amino acid-containing peptide that binds insulin is described. The amino acid sequence of the insulin-binding peptide (IBP) was determined from the strand of DNA complementary to the strand of DNA coding for the insulin molecule in the domain of the insulin monomer believed to interact with the insulin receptor. The IBP (Cys-Val-Glu-Glu-Ala-Ser) binds specifically to insulin in a saturable manner with a Kd of 3 nM. This binding process is time dependent and slightly temperature dependent, and the peptide appears to interact with insulin near the carboxyl terminus of the B-chain of insulin. Incubation of insulin with the peptide decreases insulin binding to the insulin receptor by 50%, with no effect on the affinity of insulin for the receptor and no effect on cellular insulin-stimulated deoxyglucose uptake. A polyclonal antibody produced against the IBP will inhibit specific insulin binding to intact cells by approximately 50%, with no effects on insulin-stimulated glucose uptake. From this data, we suggest that there are at least two domains of the insulin molecule through which it interacts with its receptor, the "binding region" of insulin, which is the domain blocked by the IBP, and the "message region" of insulin, through which insulin not only binds to the receptor, but also generates the cellular signal.
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis; D001483:Base Sequence; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000138:chemical synthesis* / Q000378:metabolism; D002478:Cells, Cultured; D007106:Immune Sera; D007328:Insulin / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D011972:Receptor, Insulin / Q000235:genetics / Q000378:metabolism*; D013816:Thermodynamics
chemical_list: D000936:Antigen-Antibody Complex; D002352:Carrier Proteins; D007106:Immune Sera; D007328:Insulin; D047908:Intracellular Signaling Peptides and Proteins; C057274:insulin-binding peptide; D011972:Receptor, Insulin