Subcellular distribution of proteolytic activities degrading bioactive peptides and analogues in the rat small intestinal and colonic enterocytes.
The objective of this study was to compare, in rat small intestinal and colonic enterocytes, subcellular distributions of activities degrading the large peptides, neurotensin, acetylneurotensin (8-13), GRF(1-29)NH2 (human growth hormone releasing factor fragment), (desNH2Tyr1,D-Ala2,Ala15)-GRF(1-29)NH2, insulin, and insulin B-chain. Proteolytic activities degrading individual peptides in the 10,000-g pellet, rich in intracellular organelles, 27,000-g pellet, rich in brush-border membrane, 100,000-g pellet, and 100,000-g supernatant, rich in cytosol, were determined and compared for both the small intestine and colon. In colonic fractions, the cytosol had highest activity (g protein)-1 degrading three out of four peptides tested, while in small intestinal fractions, the 27,000-g pellet had the highest activity (g protein)-1, degrading four out of five peptides tested. In both small intestine and colon, the cytosol had a higher percentage of total proteolytic activity degrading each of the above polypeptides and the highest insulin-degrading activity (g protein)-1. The results suggest that at pH 7.5, proteolytic activities (g protein)-1 in the fraction of subcellular organelles are much lower than those in cytosol and that cytosolic proteolytic activities degrading polypeptides and analogues are significant.
Bai
J P
JP
College of Pharmacy, University of Minnesota, Minneapolis 55455.
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003106:Colon / Q000378:metabolism* / Q000648:ultrastructure; D003600:Cytosol / Q000378:metabolism; D013007:Growth Hormone-Releasing Hormone / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism*; D007421:Intestine, Small / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008861:Microsomes / Q000378:metabolism; D008871:Microvilli / Q000378:metabolism; D008969:Molecular Sequence Data; D009496:Neurotensin / Q000031:analogs & derivatives / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution
D007328:Insulin; D010446:Peptide Fragments; D009496:Neurotensin; C034659:acetylneurotensin (8-13); D013007:Growth Hormone-Releasing Hormone; C059148:somatotropin releasing hormone (1-29)