Biochemical characterization of the catalytic domain of human matrix metalloproteinase 19 -: Evidence for a role as a potent basement membrane degrading enzyme

被引:119
作者
Stracke, JO [1 ]
Hutton, M
Stewart, M
Pendás, AM
Smith, B
López-Otin, C
Murphy, G
Knäuper, V
机构
[1] Univ E Anglia, Sch Biol Sci, Norwich NR4 7TJ, Norfolk, England
[2] Univ Oviedo, Dept Bioquim & Biol Mol, E-33006 Oviedo, Spain
[3] Celltech Ltd, Slough SL1 4EN, Berks, England
关键词
D O I
10.1074/jbc.275.20.14809
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have recently cloned MMP-19 a novel matrix metalloproteinase, which, due to unique structural features, was proposed to represent the first member of a new MMP subfamily (Pendas, A.M., Knauper, V., Puente, X. S., Llano, E., Mattei, M. G., Apte, S., Murphy, G., and Lopez-Otin, C. (1997) J. Biol. Chem. 272, 4281-4286). A recombinant COOH-terminal deletion mutant of MMP-19 (pro Delta(260-508)MMP-19), comprising the propeptide and the catalytic domain, was expressed in Escherichia coli, refolded, and purified. Interestingly, we found that pro Delta(260-508)MMP-19 has the tendency to autoactivate, whereby the Lys(97)-Tyr(98) peptide bond is hydrolyzed, resulting in free catalytic domain. Mutation of two residues (Glu(88), Pro and Pro(90) --> Val) within the propeptide latency motif did not prevent autoactivation but the autolysis rate was somewhat reduced. Analysis of the substrate specificity revealed that the catalytic domain of MMP-19 was able to hydrolyze the general MMP substrate Mca-Pro-Leu-Gly-Dpa-Ala-Arg-NH, and, with higher efficiency, the stromelysin substrate Mca-Pro-Leu-Ala-Nva-Dpa-Ala-Arg-NH2. Kinetic analysis of the interactions of the catalytic domain of MMP-19 with the natural MMP inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), showed strong inhibition using TIMP-2, TIMP-3, and TIMP-4, while TIMP-1 was less efficient. We also demonstrated that synthetic hydroxamic acid-based compounds efficiently inhibited the enzyme. The catalytic domain of MMP-19 was able to hydrolyze the basement membrane components type TV collagen, laminin, and nidogen, as well as the large tenascin-C isoform, fibronectin, and type I gelatin in vitro, suggesting that MMP-19 is a potent proteinase capable of hydrolyzing a broad range of extracellular matrix components. Neither the catalytic domain nor the full-length MMP-19 was able to degrade triple-helical collagen. Finally, and in contrast to studies with other MMPs, MMP-19 catalytic domain was not able to activate any of the latent MMPs tested in vitro.
引用
收藏
页码:14809 / 14816
页数:8
相关论文
共 46 条
[21]   The matrix metalloproteinase RASI-1 is expressed in synovial blood vessels of a rheumatoid arthritis patient [J].
Kolb, C ;
Mauch, S ;
Peter, HH ;
Krawinkel, U ;
Sedlacek, R .
IMMUNOLOGY LETTERS, 1997, 57 (1-3) :83-88
[22]   Analysis of 16 different matrix metalloproteinases (MMP-1 to MMP-20) in the synovial membrane:: different profiles in trauma and rheumatoid arthritis [J].
Konttinen, YT ;
Ainola, M ;
Valleala, H ;
Ma, J ;
Ida, H ;
Mandelin, J ;
Kinne, RW ;
Santavirta, S ;
Sorsa, T ;
López-Otín, C ;
Takagi, M .
ANNALS OF THE RHEUMATIC DISEASES, 1999, 58 (11) :691-697
[23]  
Lovejoy B, 1999, NAT STRUCT BIOL, V6, P217
[24]   MEMBRANE-ANCHORED GROWTH-FACTORS [J].
MASSAGUE, J ;
PANDIELLA, A .
ANNUAL REVIEW OF BIOCHEMISTRY, 1993, 62 :515-541
[25]   Structural and genetic analysis of laminin-nidogen interaction [J].
Mayer, U ;
Kohfeldt, E ;
Timpl, R .
MORPHOGENESIS: CELLULAR INTERACTIONS, 1998, 857 :130-142
[26]   Residue 2 of TIMP-1 is a major determinant of affinity and specificity for matrix metalloproteinases but effects of substitutions do not correlate with those of the corresponding P1′ residue of substrate [J].
Meng, Q ;
Malinovskii, V ;
Huang, W ;
Hu, YJ ;
Chung, L ;
Nagase, H ;
Bode, W ;
Maskos, K ;
Brew, K .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (15) :10184-10189
[27]   MATRIX METALLOPROTEINASE DEGRADATION OF ELASTIN, TYPE-IV COLLAGEN AND PROTEOGLYCAN - A QUANTITATIVE COMPARISON OF THE ACTIVITIES OF 95 KDA AND 72 KDA GELATINASES, STROMELYSIN-1 AND STROMELYSIN-2 AND PUNCTUATED METALLOPROTEINASE (PUMP) [J].
MURPHY, G ;
COCKETT, MI ;
WARD, RV ;
DOCHERTY, AJP .
BIOCHEMICAL JOURNAL, 1991, 277 :277-279
[28]   THE N-TERMINAL DOMAIN OF TISSUE INHIBITOR OF METALLOPROTEINASES RETAINS METALLOPROTEINASE INHIBITORY ACTIVITY [J].
MURPHY, G ;
HOUBRECHTS, A ;
COCKETT, MI ;
WILLIAMSON, RA ;
OSHEA, M ;
DOCHERTY, AJP .
BIOCHEMISTRY, 1991, 30 (33) :8097-8101
[29]   High resolution structure of the N-terminal domain of tissue inhibitor of metalloproteinases-2 and characterization of its interaction site with matrix metalloproteinase-3 [J].
Muskett, FW ;
Frenkiel, TA ;
Feeney, J ;
Freedman, RB ;
Carr, MD ;
Williamson, RA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (34) :21736-21743
[30]   Involvement of a region near valine-69 of tissue inhibitor of metalloproteinases (TIMP)-1 in the interaction with matrix metalloproteinase 3 (stromelysin 1) [J].
Nagase, H ;
Suzuki, K ;
Cawston, TE ;
Brew, K .
BIOCHEMICAL JOURNAL, 1997, 325 :163-167