Cholecystokinin octa- and tetrapeptide degradation by synaptic membranes. I. Evidence for competition with enkephalins for in vitro common degradation pathways.
Degradation of CCK-4 and -8 by purified synaptic membranes was followed by measuring the fluorescence of tryptophan released from the peptides after separation of degradation products by HPLC. For enkephalins and related fragments, the release of tyrosine was monitored using the same method. Kinetics of hydrolysis of CCK-like peptides indicated a rapid processing of CCK-4 and a slower breakdown of CCK-8 (with a greater resistance of the sulfated form of CCK-8 as compared to the unsulfated peptide). Leu- and met-enkephalins were degraded at the same rate while their N-terminal tri- and dipeptides were hydrolysed more slowly. When CCK-4 or CCK-8 were incubated in the presence of leu-enkephalin, a dose-dependent inhibition of the release of tryptophan was observed. Enkaphalin fragments do not modify the kinetics of degradation of CCK-4. The degradation of leu-enkephalin was inhibited in a dose-dependent manner by the presence of CCK-related peptides in the medium. After solubilization of membrane-bound enzymes by Triton X-100 followed by chromatography on DEAE cellulose, five peaks of CCK-4 degrading activity were detected (two minor and three major peaks). With enkephalin as substrate, five peaks were also observed; the three major activities were the same as those detected for CCK-4.
D000626:Aminopeptidases / Q000037:antagonists & inhibitors; D000818:Animals; D002766:Cholecystokinin / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D002848:Chromatography, DEAE-Cellulose; D004723:Endorphins / Q000378:metabolism*; D004743:Enkephalin, Leucine; D004745:Enkephalins / Q000378:metabolism* / Q000494:pharmacology; D005755:Gastrins / Q000378:metabolism*; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D010446:Peptide Fragments / Q000378:metabolism*; D051381:Rats; D012844:Sincalide; D013570:Synaptic Membranes / Q000187:drug effects / Q000378:metabolism*; D013758:Tetragastrin / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism
D004723:Endorphins; D004745:Enkephalins; D005755:Gastrins; D010446:Peptide Fragments; D013758:Tetragastrin; D004743:Enkephalin, Leucine; D014364:Tryptophan; D002766:Cholecystokinin; D000626:Aminopeptidases; D007930:Leucine; C012211:ubenimex; D012844:Sincalide