title: A 3,4-dichloroisocoumarin-resistant component of the multicatalytic proteinase complex.
abstract: The multicatalytic proteinase complex (MPC) exhibits three proteolytic activities designated as trypsin-like, chymotrypsin-like, and peptidylglutamyl-peptide hydrolyzing (PGPHA). Evidence based on inhibitor and specificity studies indicates that each of the three activities is associated with a different component of the complex. Inactivation of the three activities by the serine proteinase inhibitor, 3,4-dichloroisocoumarin (DCI), reveals the presence of an additional DCI-resistant component that cleaves natural peptides including neurotensin, dynorphin, angiotensin II, the oxidized B-chain of insulin, and also proinsulin at a rate greater than that of the native uninhibited complex. Examination of the reaction products of neurotensin (NT) and proinsulin degradation showed cleavage of the Ile12-Leu13 bond in NT and cleavage of the Leu44-Ala45 and Val39-Gly40 bonds within the connecting peptide (C-chain) of bovine proinsulin, suggesting preferential cleavage of bonds on the carboxyl side of branched chain amino acids. Although resistant to inhibition by DCI, the component was sensitive to inhibition by the isocoumarin derivatives, 7-amino-4-chloro-3-[3-(isothioureido)propoxy]isocoumarin and 4-chloro-7-guanidino-3-(2-phenylethoxy)isocoumarin. Degradation of NT was activated by leupeptin, chymostatin, and antipain indicating that binding of these aldehyde inhibitors at one site can stimulate proteolytic activity at a different site of the complex. The DCI-resistant component seems to constitute a major component of the complex active in degradation of natural peptides and proteins.
authors.affiliation: Department of Pharmacology, Mount Sinai School of Medicine, City University of New York, New York 10029.
authors.forename: C
authors.initials: C
authors.lastname: Cardozo
authors.forename: A
authors.initials: A
authors.lastname: Vinitsky
authors.forename: M C
authors.initials: MC
authors.lastname: Hidalgo
authors.forename: C
authors.initials: C
authors.lastname: Michaud
authors.forename: M
authors.initials: M
authors.lastname: Orlowski
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003374:Coumarins / Q000494:pharmacology*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004351:Drug Resistance; D049934:Isocoumarins; D007700:Kinetics; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000378:metabolism*; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism; D010902:Pituitary Gland / Q000201:enzymology; D011480:Protease Inhibitors / Q000494:pharmacology*; D046988:Proteasome Endopeptidase Complex; D013379:Substrate Specificity
chemical_list: D003374:Coumarins; D049934:Isocoumarins; D009097:Multienzyme Complexes; D009842:Oligopeptides; D011480:Protease Inhibitors; C046235:3,4-dichloroisocoumarin; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex