Transpeptidation reactions of porcine pepsin. Formation of tetrapeptides from dipeptide substrates.
Pepsin-catalyzed transpeptidation was studied by high resolution 75 MHz 13C nuclear magnetic resonance spectroscopy. Enrichment with 13C at the carbonyl carbons of the substrates Leu-Tyr-NH2 and Leu-Leu-NH2 facilitated detection and identification of the transpeptidation and hydrolysis products of enzymic action. Porcine pepsin was found in each case to synthesize and release the tetrapeptide Leu-Leu-Leu-Leu as the primary product of transpeptidation, the longest oligomeric product of transpeptidation observed to date. Productive binding of the dipeptide substrates into the active site groove of pepsin required an induction period of several minutes. Quenching experiments suggested the presence of strongly bound intermediate forms of Leu and Leu-Leu prior to observation of any enzyme-free products. The finding of the tetrapeptide as a primary product is discussed as an instance where transpeptidation of the tripeptide competes successfully with the action of pepsin subsite S3 as a trigger for product release.
D000818:Animals; D004151:Dipeptides / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007930:Leucine; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D009842:Oligopeptides / Q000378:metabolism*; D010434:Pepsin A / Q000378:metabolism*; D013552:Swine
D004151:Dipeptides; D009842:Oligopeptides; D010434:Pepsin A; D007930:Leucine