Differential inhibition of membrane type 3 (MT3)-matrix metalloproteinase (MMP) and MT1-MMP by tissue inhibitor of metalloproteinase (TIMP)-2 and TIMP-3 rgulates pro-MMP-2 activation.
The membrane type (MT)-matrix metalloproteinases (MMPs) constitute a subgroup of membrane-anchored MMPs that are major mediators of pericellular proteolysis and physiological activators of pro-MMP-2. The MT-MMPs also exhibit differential inhibition by members of the tissue inhibitor of metalloproteinase (TIMP) family. Here we investigated the processing, catalytic activity, and TIMP inhibition of MT3-MMP (MMP-16). Inhibitor profile and mutant enzyme studies indicated that MT3-MMP is regulated on the cell surface by autocatalytic processing and ectodomain shedding. Inhibition kinetic studies showed that TIMP-3 is a high affinity inhibitor of MT3-MMP when compared with MT1-MMP (K(i) = 0.008 nm for MT3-MMP versus K(i) = 0.16 nm for MT1-MMP). In contrast, TIMP-2 is a better inhibitor of MT1-MMP. MT3-MMP requires TIMP-2 to accomplish full pro-MMP-2 activation and this process is enhanced in marimastatpretreated cells, consistent with regulation of active enzyme turnover by synthetic MMP inhibitors. TIMP-3 also enhances the activation of pro-MMP-2 by MT3-MMP but not by MT1-MMP. TIMP-4, in contrast, cannot support pro-MMP-2 activation with either enzyme. Affinity chromatography experiments demonstrated that pro-MMP-2 can assemble trimolecular complexes with a catalytic domain of MT3-MMP and TIMP-2 or TIMP-3 suggesting that pro-MMP-2 activation by MT3-MMP involves ternary complex formation on the cell surface. These results demonstrate that TIMP-3 is a major regulator of MT3-MMP activity and further underscores the unique interactions of TIMPs with MT-MMPs in the control of pericellular proteolysis.
Zhao
Huiren
H
Department of Pathology, School of Medicine, Wayne State University, Detroit, Michigan 48201, USA.
Bernardo
M Margarida
MM
Osenkowski
Pamela
P
Sohail
Anjum
A
Pei
Duanqing
D
Nagase
Hideaki
H
Kashiwagi
Masahide
M
Soloway
Paul D
PD
DeClerck
Yves A
YA
Fridman
Rafael
R
D000818:Animals; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000235:genetics; D004792:Enzyme Precursors / Q000096:biosynthesis; D015971:Gene Expression Regulation, Enzymologic; D006868:Hydrolysis; D053511:Matrix Metalloproteinase 14; D053513:Matrix Metalloproteinase 16; D019278:Matrix Metalloproteinase 3 / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000235:genetics; D000090023:Metallothionein 3; D051379:Mice; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism*; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics / Q000378:metabolism*
D004792:Enzyme Precursors; D061965:Matrix Metalloproteinase Inhibitors; D000090023:Metallothionein 3; C507096:Mmp14 protein, mouse; C507104:Mmp16 protein, mouse; C000715430:Mt3 protein, mouse; D019717:Tissue Inhibitor of Metalloproteinase-3; D019716:Tissue Inhibitor of Metalloproteinase-2; D053513:Matrix Metalloproteinase 16; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D053511:Matrix Metalloproteinase 14