Characterization of Rat Uterine Matrilysin and Its cDNA
RELATIONSHIP TO HUMAN PUMP-1 AND ACTIVATION OF PROCOLLAGENASES (*)
- Susan R. Abramson(4)(1)(§),
- Gregory E. Conner(2),
- Hideaki Nagase(5),
- Isaac Neuhaus(1) and
- J. Frederick Woessner, Jr.(1)(3)
- From the (1)Departments of Biochemistry and Molecular Biology,
- (2)Cell Biology and Anatomy, and
- (3)Medicine, University of Miami School of Medicine, Miami, Florida 33136, the
- (4)Division of Research, Cleveland Clinic Florida, Ft. Lauderdale, Florida 33309, and the
- (5)Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, Kansas 66160
- § Supported by National Institutes of Health Training Grant HD-07129 and a University of Miami Graduate Fellowship. To whom correspondence should be addressed: Cleveland Clinic Florida, Div. of Research, 3000 W. Cypress Creek Rd., Ft. Lauderdale, FL 33309. 305-978-5898; Fax: 305-978-5006.
Abstract
A small uterine metalloproteinase of the rat has been shown by amino acid and cDNA sequencing to be orthologous to human pump-1.
Both proteinases are now designated as matrilysin or matrix metalloproteinase 7. The properties of purified uterine metalloproteinase
and recombinant pump-1 were compared. Their specificities on substrates (gelatins, fibronectin, transferrin, elastin, Azocoll,
and (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3,[2, 4-dinitrophenyl]-L-2, 3-diaminopropionyl)-Ala-Arg-NH2) are similar and distinct from those of the stromelysins and gelatinases. The two matrilysins have similar sensitivity to
hydroxamate and pseudopeptide inhibitors. Rat matrilysin selectively cleaves the α2(I) chain of rat gelatin, producing major
cuts at Gly
-
-Ile
, Gly
-
-Leu
, and Gly
-
-Ile
. Rat matrilysin produces maximum activation of latent human interstitial collagenase 1 (pro-matrix metalloproteinase 1) when
added in the presence of 4-aminophenylmercuric acetate (APMA) by cleaving the Gln
-
-Phe
bond. Rat and human matrilysin do not directly activate latent rat collagenase 3 (matrix metalloproteinase 13) and do not
enhance its activation when added together with APMA. Autoactivation of collagenase 3 in the presence of APMA results in cleavage
at Val
-
-Tyr
corresponding to the Gln
-
-Phe
cleavage in collagenase 1. Thus collagenase 3 is capable of maximal autoactivation, whereas collagenase 1 is dependent upon
another matrix metalloproteinase in order to be activated to its full potential.
Footnotes
-
↵* This investigation was supported by National Institutes of Health Grants HD-06773 (to J. F. W.), AR-39189 (to H. N.), and GM-35812 (to G. E. C.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
-
↵1 The abbreviations used are:
- DNP-peptide
-
DNP-Pro-Leu-Gly-Ile-Ala-Gly-Glu-D-Arg
- APMA
-
4-aminophenylmercuric acetate
- Mca-peptide
-
(7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3,[2, 4-dinitrophenyl]-L-2, 3-diaminopropionyl)-Ala-Arg-NH2
- PAGE
-
polyacrylamide electrophoresis
- BB94
-
[4-(N-hydroxyamino)-2R-isobutyl-3S-(thiopen-2-ylthiomethyl)-succinyl]-L-phenylalanine-N-methylamide
- SC 40827
-
N-[3-N-(benzyloxycarbonyl)amino-1-(R)carboxypropyl]-L-leucyl-O-methyl-L-tyrosine-N-methylamide
- SC 44463
-
N4-hydroxy-N1-{1S-[(4-methoxphenyl)methyl]-2-(methylamino)-2-oxoethyl}-2R-(2-methylpro-pyl)butane-diamide
- PCR
-
polymerase chain reaction.
-
↵2 The cDNA sequence has been deposited in GenBank
, accession number L24374.
-
↵3 K. Appelt, personal communication.
