Conformational and biological properties of glucagon fragments containing residues 1-17 and 19-29.
The 29 amino acid polypeptide hormone glucagon was cleaved into two large fragments by the enzyme clostripain. The conformational properties of these two fragments were monitored by circular dichroism at pH 2 and 12 in both the presence and absence of sodium dodecyl sulfate. Both glucagon (1-17) and glucagon (19-29) have reduced abilities to fold in aqueous solution. However, both fragments can take on structure of higher apparent helical content in acidic solution in the presence of sodium dodecyl sulfate but only the glucagon (19-29) retains this conformation at high pH. Neither of the two fragments react with dimyristoylphosphatidylcholine as the intact peptide does. Only the carboxyl terminal fragment was capable of reacting with an antibody specific for glucagon. The glucagon (1-17) has markedly reduced affinity for binding to the glucagon receptor as well as markedly reduced ability to stimulate adenylate cyclase activity which is not affected by the presence of glucagon (19-29). It is proposed that the intact sequence provides specific groups required for activity as well as the potential for forming a stable amphipathic helix, both of which are necessary for full biological activity at low peptide concentrations.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D003546:Cysteine Endopeptidases; D004134:Dimyristoylphosphatidylcholine; D010450:Endopeptidases; D005934:Glucagon / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D008099:Liver / Q000378:metabolism; D010446:Peptide Fragments / Q000302:isolation & purification* / Q000494:pharmacology; D011487:Protein Conformation; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism; D018027:Receptors, Glucagon; D013329:Structure-Activity Relationship; D013552:Swine
D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D018027:Receptors, Glucagon; C039265:glucagon (19-29); C039264:glucagon (1-17); D005934:Glucagon; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; C019913:clostripain; D000262:Adenylyl Cyclases; D004134:Dimyristoylphosphatidylcholine