The matrix metalloproteinase pump-1 catalyzes formation of low molecular weight (pro)urokinase in cultures of normal human kidney cells

The enzyme responsible for the metalloproteinase activity which cleaves the Glu143-Leu144 bond of (pro)urokinase has been isolated from the conditioned medium of cultured normal human kidney cells. Using S-Sepharose and Cibacron Blue-agarose chromatography, then C-4 reversed phase high pressure liqu...

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Bibliographic Details
Published in:	The Journal of biological chemistry Vol. 267; no. 20; pp. 13803 - 13806
Main Authors:	MARCOTTE, P. A, KOZAN, I. M, DORWIN, S. A, RYAN, J. M
Format:	Journal Article
Language:	English
Published:	Bethesda, MD American Society for Biochemistry and Molecular Biology 15-07-1992
Subjects:	Amino Acid Sequence Analytical, structural and metabolic biochemistry Biological and medical sciences cells Cells, Cultured Chromatography, High Pressure Liquid Electrophoresis, Polyacrylamide Gel enzymatic activity Enzyme Precursors - isolation & purification Enzyme Precursors - metabolism Enzymes and enzyme inhibitors formation Fundamental and applied biological sciences. Psychology Humans Hydrolases Kidney - enzymology man Matrix Metalloproteinase 7 Metalloendopeptidases - genetics Metalloendopeptidases - isolation & purification Metalloendopeptidases - metabolism metalloproteinase PUMP-1 Molecular Sequence Data Molecular Weight plasminogen activator precursors purification Urokinase-Type Plasminogen Activator - isolation & purification Urokinase-Type Plasminogen Activator - metabolism Human Urokinase Purification Enzyme Affinity chromatography Substrate specificity HPLC chromatography Metalloproteinase Kidney
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Summary:	The enzyme responsible for the metalloproteinase activity which cleaves the Glu143-Leu144 bond of (pro)urokinase has been isolated from the conditioned medium of cultured normal human kidney cells. Using S-Sepharose and Cibacron Blue-agarose chromatography, then C-4 reversed phase high pressure liquid chromatography, a protein of about 20,000 Da was isolated. Through an identical amino-terminal sequence, the protein was shown to be the matrix metalloproteinase previously referred to in the literature as "pump-1" (putative metalloproteinase). When aprotinin was added during the course of the purification, the major species isolated was the zymogen form (28,000 Da) of pump-1. Pump-1 has been shown to efficiently cleave the susceptible bond of both pro-urokinase (single-chain) and active (two-chain) urokinase and thereby produce the corresponding low molecular weight forms. The amino-terminal sequences of the A and B chains of low molecular weight urokinase prepared by action of pump-1 on recombinant high molecular weight urokinase are identical to those of the low molecular weight urokinase isolated from human kidney cell culture. Since the reaction of urokinase with this metalloproteinase results in separation of its serine proteinase region from the domain which mediates binding to the urokinase receptor, it may be of importance in the regulation of the functional activity of the plasminogen activator in cellular processes.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0021-9258 1083-351X
DOI:	10.1016/S0021-9258(19)49637-3