The unregulated activities of matrix metalloproteinases (MMPs) are implicated in disease processes including arthritis and tumor cell invasion and metastasis. MMP activities are controlled by four homologous endogenous protein inhibitors, tissue inhibitors of metalloproteinases (TIMPs), yet different TIMPs show little specificity for individual MMPs. The large interaction interface in the TIMP-1·MMP-3 complex includes a contiguous region of TIMP-1 around the disulfide bond between Cys1 and Cys70 that inserts into the active site of MMP-3. The effects of fifteen different substitutions for threonine 2 of this region reveal that this residue makes a large contribution to the stability of complexes with MMPs and has a dominant influence on the specificity for different MMPs. The size, charge, and hydrophobicity of residue 2 are key factors in the specificity of TIMP. Threonine 2 of TIMP-1 interacts with the S1′ specificity pocket of MMP-3, which is a key to substrate specificity, but the structural requirements in TIMP-1 residue 2 for MMP binding differ greatly from those for the corresponding residue of a peptide substrate. These results demonstrate that TIMP variants with substitutions for Thr2 represent suitable starting points for generating more targeted TIMPs for investigation and for intervention in MMP-related diseases. The unregulated activities of matrix metalloproteinases (MMPs) are implicated in disease processes including arthritis and tumor cell invasion and metastasis. MMP activities are controlled by four homologous endogenous protein inhibitors, tissue inhibitors of metalloproteinases (TIMPs), yet different TIMPs show little specificity for individual MMPs. The large interaction interface in the TIMP-1·MMP-3 complex includes a contiguous region of TIMP-1 around the disulfide bond between Cys1 and Cys70 that inserts into the active site of MMP-3. The effects of fifteen different substitutions for threonine 2 of this region reveal that this residue makes a large contribution to the stability of complexes with MMPs and has a dominant influence on the specificity for different MMPs. The size, charge, and hydrophobicity of residue 2 are key factors in the specificity of TIMP. Threonine 2 of TIMP-1 interacts with the S1′ specificity pocket of MMP-3, which is a key to substrate specificity, but the structural requirements in TIMP-1 residue 2 for MMP binding differ greatly from those for the corresponding residue of a peptide substrate. These results demonstrate that TIMP variants with substitutions for Thr2 represent suitable starting points for generating more targeted TIMPs for investigation and for intervention in MMP-related diseases. matrix metalloproteinases tissue inhibitors of metalloproteinases N-terminal domain of tissue inhibitor of metalloproteinases-1 The matrix metalloproteinases (MMPs)1 are a family of about twenty Zn2+-dependent endopeptidases that have important roles in connective tissue turnover during physiological processes including development, morphogenesis, and wound healing (1Woessner Jr., J.F. Ann. N. Y. Acad. Sci. 1994; 732: 11-21Crossref PubMed Scopus (436) Google Scholar,2Nagase H. Hooper N.M. Zinc Metalloproteinases in Health and Disease. Taylor & Francis Ltd., London1996: 153-204Google Scholar). Their activities in the extracellular matrix are stringently regulated through transcriptional control, zymogen activation, and the actions of four endogenous inhibitory proteins, tissue inhibitors of metalloproteinases (TIMPs) 1 to 4 (3Docherty A.J.P. Lyons A. Smith B.J. Wright E.M. Stephens P.E. Harris T.J.R. Murphy G. Reynolds J.J. Nature. 1985; 318: 66-69Crossref PubMed Scopus (582) Google Scholar, 4Boone T.C. Johnson M.J. DeClerck Y.A. Langley K.E. Proc. Natl. Acad. Sci. U. S. A. 1990; 87: 2800-2804Crossref PubMed Scopus (180) Google Scholar, 5Pavloff N. Staskus P.W. Kishnani N.S. Hawkes S.P. J. Biol. Chem. 1992; 267: 17321-17326Abstract Full Text PDF PubMed Google Scholar, 6Silbiger S.M. Jacobsen V.L. Cupples R.L. Koski R.A. Gene. 1994; 141: 293-297Crossref PubMed Scopus (66) Google Scholar, 7Greene J. Wang M. Liu Y.E. Raymond L.A. Rosen C. Shi Y.E. J. Biol. Chem. 1996; 271: 30375-30380Abstract Full Text Full Text PDF PubMed Scopus (475) Google Scholar). Normal matrix homeostasis is associated with an appropriate balance between the levels of TIMPs and active MMPs, whereas an imbalance involving excess MMP activity is linked with disease processes including arthritis, tumor cell metastasis, and tissue invasion and atherosclerosis (1Woessner Jr., J.F. Ann. N. Y. Acad. Sci. 1994; 732: 11-21Crossref PubMed Scopus (436) Google Scholar, 2Nagase H. Hooper N.M. Zinc Metalloproteinases in Health and Disease. Taylor & Francis Ltd., London1996: 153-204Google Scholar).Mammalian TIMPs have an N-terminal domain of about 125 amino acids and a smaller C-terminal domain of about 65 amino acids; each domain is stabilized by three disulfide bonds (8Williamson E.A. Marston F.A.O. Angal S. Koklitis P. Panico M. Morris H.R. Carne A.F. Smith B.J. Harris T.J.R. Freedman R.B. Biochem. J. 1990; 268: 267-274Crossref PubMed Scopus (155) Google Scholar). The N-terminal domains of different TIMPs fold into a correct native structure which carries the inhibitory activity against MMPs (9Murphy G. Houbrechts A. Cockett M.I. Williamson R.O.-A. Shea M. Docherty A.J.P. Biochemistry. 1991; 30: 8097-8102Crossref PubMed Scopus (283) Google Scholar, 10O'Shea M. Willenbrock F. Williamson R.A. Cockett M.I. Freedman R.B. Reynolds J.J. Docherty A.J.P. Murphy G. Biochemistry. 1992; 31: 10146-10152Crossref PubMed Scopus (73) Google Scholar, 11Huang W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar). Although correctly folded and functional C-terminal domains have not been described, truncation experiments indicate that this region is responsible for the interactions of TIMPs with pro-MMPs (12Murphy G. Willenbrock F. Methods Enzymol. 1995; 248: 496-510Crossref PubMed Scopus (243) Google Scholar, 13Bigg H.F. Shi Y.E. Liu Y.E. Steffensen B. Overall C.M. J. Biol. Chem. 1997; 272: 15496-15500Crossref PubMed Scopus (143) Google Scholar). There is little specificity in the inhibitory actions of TIMPs on metalloproteinases, with the exception of the ability of TIMP-2 and TIMP-3 to inhibit membrane-type metalloproteinases-1 and -2, whereas TIMP-1 is a poor inhibitor of these enzymes (12Murphy G. Willenbrock F. Methods Enzymol. 1995; 248: 496-510Crossref PubMed Scopus (243) Google Scholar, 13Bigg H.F. Shi Y.E. Liu Y.E. Steffensen B. Overall C.M. J. Biol. Chem. 1997; 272: 15496-15500Crossref PubMed Scopus (143) Google Scholar, 14Will H. Atkinson S.J. Butler G.S. Smith B. Murphy G. J. Biol. Chem. 1996; 271: 17119-17123Abstract Full Text Full Text PDF PubMed Scopus (506) Google Scholar). However, the interactions of TIMPs with pro-MMPs are more specific. For example, TIMP-2 and TIMP-4 form specific complexes with pro-MMP-2 whereas TIMP-1 to to activities MMP inhibitors and in binding to TIMPs the of of in tissue K. K. K. FEBS Lett. 1992; PubMed Scopus Google Scholar, K. A. J. Sci. 1994; PubMed Google and have activity B. K. B. Murphy G. S. B. Sci. 1997; Google Scholar). However, the structural of these activities is have been for a complex of TIMP-1 with the domain of K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google and a complex of TIMP-2 with the domain of membrane-type matrix 1 C. W. J.J. A. H. K. J. PubMed Scopus Google Scholar). with a structure of the N-terminal domain of R.A. G. Murphy G. Docherty A.J.P. Freedman R.B. J. Biochemistry. 1994; PubMed Scopus Google Scholar, J. Freedman R.B. Williamson R.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google these reveal that the inhibitory domain of TIMP of a with three associated the of of the protein family J. PubMed Scopus Google Scholar). The TIMP-1·MMP-3 structure that the interactions between TIMP and the the N-terminal and of the between and interactions are through the and in the C-terminal domain 1 of are by to the disulfide bond between Cys1 and and K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). The N-terminal Cys1 is a key to the inhibitory of TIMP on of the of the and the through the and peptide 1 are in the are in the of the and in the of interactions involving of the structure C. W. J.J. A. H. K. J. PubMed Scopus Google site of TIMP by this structure is with the results of a which that TIMP-1 activity is the peptide bond is by but that this is by complex between TIMP-1 and H. Suzuki K. Brew K. Biochem. J. 1997; PubMed Scopus Google Scholar). of show that substitutions a of between and have effects on the for MMP-3, whereas that the Cys1 to Cys70 disulfide and substitutions for large effects on activity W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google Scholar). whereas substitutions that activity have effects on binding to and MMP-3, the of for Thr2 a in binding to to W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google Scholar). is in with the which indicate that Thr2 of TIMP-1 and of TIMP-2 with the region of the metalloproteinases that to the binding site for the residue of peptide the residue that has a dominant in MMP specificity K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar, C. W. J.J. A. H. K. J. PubMed Scopus Google of a of the structural of TIMP-1 specificity and a generating variants that are more MMP inhibitors, have fifteen with substitutions for The results show that this residue has a influence on the specificity of TIMP for different metalloproteinases but show that is little between the of an amino 2 in TIMP-1 and the residue the site of a peptide substrate on activities with an MMP inhibitor substrate. The structural of these is results that protein of TIMP is a for generating more specific protein inhibitors of MMPs for of the roles of different MMPs and to the of for linked with excess activity of specific MMPs. The of targeted variants substitutions but the inhibitory of the and and TIMP-1 W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar, W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google indicate that residue 2 has a influence on specificity of the site The effects of substitutions for Thr2 about the of MMP binding by and TIMP variants with that the inhibitors for and MMP-3, on and have different substitutions The of for Thr2 a protein that is an inhibitor yet a native structure by Although the of TIMP not of the in TIMP-1 a protein that to the interactions of the C-terminal domain in a that has little for a active site and to the of TIMP is with cell and activity K. K. K. FEBS Lett. 1992; PubMed Scopus Google Scholar, K. A. J. Sci. 1994; PubMed Google Scholar, B. K. B. Murphy G. S. B. Sci. 1997; Google of the of residue 2 with the for different MMPs is to the structural of the effects of on the TIMP-1·MMP-3 the N-terminal region of has a different structure from that in K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). on the structure of that in TIMP during complex J. Freedman R.B. Williamson R.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). in the site the interaction with MMPs through effects on to different MMPs by from the disulfide The of for the that interactions of the S1′ pocket more of the binding for the interaction with with the structure of the S1′ pocket of is and those of MMPs. which in and MMP-3, into the S1′ pocket the in a more pocket Smith B. Biol. 1994; PubMed Scopus Google Scholar). The for the of for and threonine for example, and is a in the binding of to The specificity pocket of an MMP with substrate inhibitor to that the binding of TIMP by the ability of the residue 2 to with of in the of complexes S. Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar). has been that of the of each is on of the TIMP-1·MMP-3 complex K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). The C-terminal domain of TIMP-1 for of these and of the of interaction W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar). The results show that a residue of the site of a large influence on the and specificity of binding to MMPs. The of this is the of Thr2 from to on complex with the for H. Hooper N.M. Zinc Metalloproteinases in Health and Disease. Taylor & Francis Ltd., London1996: 153-204Google of the of the interaction site S. J. C. J. Biol. PubMed Scopus Google Scholar). The of of the of binding on of the Thr2 by the of that residue 2 of TIMP-1 a in the interaction a residue that has a large influence on the of the interaction 1995; 267: PubMed Scopus Google Scholar, Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar). form with in the structure K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google but of and and the of in that these are important for binding Thr2 W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google Scholar). the results show that the of the in the structure of the TIMP-1·MMP-3 complex are an important the structural of the inhibitory and specificity in but is important to to the stability of the The matrix metalloproteinases (MMPs)1 are a family of about twenty Zn2+-dependent endopeptidases that have important roles in connective tissue turnover during physiological processes including development, morphogenesis, and wound healing (1Woessner Jr., J.F. Ann. N. Y. Acad. Sci. 1994; 732: 11-21Crossref PubMed Scopus (436) Google Scholar,2Nagase H. Hooper N.M. Zinc Metalloproteinases in Health and Disease. Taylor & Francis Ltd., London1996: 153-204Google Scholar). Their activities in the extracellular matrix are stringently regulated through transcriptional control, zymogen activation, and the actions of four endogenous inhibitory proteins, tissue inhibitors of metalloproteinases (TIMPs) 1 to 4 (3Docherty A.J.P. Lyons A. Smith B.J. Wright E.M. Stephens P.E. Harris T.J.R. Murphy G. Reynolds J.J. Nature. 1985; 318: 66-69Crossref PubMed Scopus (582) Google Scholar, 4Boone T.C. Johnson M.J. DeClerck Y.A. Langley K.E. Proc. Natl. Acad. Sci. U. S. A. 1990; 87: 2800-2804Crossref PubMed Scopus (180) Google Scholar, 5Pavloff N. Staskus P.W. Kishnani N.S. Hawkes S.P. J. Biol. Chem. 1992; 267: 17321-17326Abstract Full Text PDF PubMed Google Scholar, 6Silbiger S.M. Jacobsen V.L. Cupples R.L. Koski R.A. Gene. 1994; 141: 293-297Crossref PubMed Scopus (66) Google Scholar, 7Greene J. Wang M. Liu Y.E. Raymond L.A. Rosen C. Shi Y.E. J. Biol. Chem. 1996; 271: 30375-30380Abstract Full Text Full Text PDF PubMed Scopus (475) Google Scholar). Normal matrix homeostasis is associated with an appropriate balance between the levels of TIMPs and active MMPs, whereas an imbalance involving excess MMP activity is linked with disease processes including arthritis, tumor cell metastasis, and tissue invasion and atherosclerosis (1Woessner Jr., J.F. Ann. N. Y. Acad. Sci. 1994; 732: 11-21Crossref PubMed Scopus (436) Google Scholar, 2Nagase H. Hooper N.M. Zinc Metalloproteinases in Health and Disease. Taylor & Francis Ltd., London1996: 153-204Google Scholar). TIMPs have an N-terminal domain of about 125 amino acids and a smaller C-terminal domain of about 65 amino acids; each domain is stabilized by three disulfide bonds (8Williamson E.A. Marston F.A.O. Angal S. Koklitis P. Panico M. Morris H.R. Carne A.F. Smith B.J. Harris T.J.R. Freedman R.B. Biochem. J. 1990; 268: 267-274Crossref PubMed Scopus (155) Google Scholar). The N-terminal domains of different TIMPs fold into a correct native structure which carries the inhibitory activity against MMPs (9Murphy G. Houbrechts A. Cockett M.I. Williamson R.O.-A. Shea M. Docherty A.J.P. Biochemistry. 1991; 30: 8097-8102Crossref PubMed Scopus (283) Google Scholar, 10O'Shea M. Willenbrock F. Williamson R.A. Cockett M.I. Freedman R.B. Reynolds J.J. Docherty A.J.P. Murphy G. Biochemistry. 1992; 31: 10146-10152Crossref PubMed Scopus (73) Google Scholar, 11Huang W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar). Although correctly folded and functional C-terminal domains have not been described, truncation experiments indicate that this region is responsible for the interactions of TIMPs with pro-MMPs (12Murphy G. Willenbrock F. Methods Enzymol. 1995; 248: 496-510Crossref PubMed Scopus (243) Google Scholar, 13Bigg H.F. Shi Y.E. Liu Y.E. Steffensen B. Overall C.M. J. Biol. Chem. 1997; 272: 15496-15500Crossref PubMed Scopus (143) Google Scholar). There is little specificity in the inhibitory actions of TIMPs on metalloproteinases, with the exception of the ability of TIMP-2 and TIMP-3 to inhibit membrane-type metalloproteinases-1 and -2, whereas TIMP-1 is a poor inhibitor of these enzymes (12Murphy G. Willenbrock F. Methods Enzymol. 1995; 248: 496-510Crossref PubMed Scopus (243) Google Scholar, 13Bigg H.F. Shi Y.E. Liu Y.E. Steffensen B. Overall C.M. J. Biol. Chem. 1997; 272: 15496-15500Crossref PubMed Scopus (143) Google Scholar, 14Will H. Atkinson S.J. Butler G.S. Smith B. Murphy G. J. Biol. Chem. 1996; 271: 17119-17123Abstract Full Text Full Text PDF PubMed Scopus (506) Google Scholar). However, the interactions of TIMPs with pro-MMPs are more specific. For example, TIMP-2 and TIMP-4 form specific complexes with pro-MMP-2 whereas TIMP-1 to to activities MMP inhibitors and in binding to TIMPs the of of in tissue K. K. K. FEBS Lett. 1992; PubMed Scopus Google Scholar, K. A. J. Sci. 1994; PubMed Google and have activity B. K. B. Murphy G. S. B. Sci. 1997; Google Scholar). However, the structural of these activities is have been for a complex of TIMP-1 with the domain of K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google and a complex of TIMP-2 with the domain of membrane-type matrix 1 C. W. J.J. A. H. K. J. PubMed Scopus Google Scholar). with a structure of the N-terminal domain of R.A. G. Murphy G. Docherty A.J.P. Freedman R.B. J. Biochemistry. 1994; PubMed Scopus Google Scholar, J. Freedman R.B. Williamson R.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google these reveal that the inhibitory domain of TIMP of a with three associated the of of the protein family J. PubMed Scopus Google Scholar). The TIMP-1·MMP-3 structure that the interactions between TIMP and the the N-terminal and of the between and interactions are through the and in the C-terminal domain 1 of are by to the disulfide bond between Cys1 and and K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). The N-terminal Cys1 is a key to the inhibitory of TIMP on of the of the and the through the and peptide 1 are in the are in the of the and in the of interactions involving of the structure C. W. J.J. A. H. K. J. PubMed Scopus Google Scholar). The site of TIMP by this structure is with the results of a which that TIMP-1 activity is the peptide bond is by but that this is by complex between TIMP-1 and H. Suzuki K. Brew K. Biochem. J. 1997; PubMed Scopus Google Scholar). of show that substitutions a of between and have effects on the for MMP-3, whereas that the Cys1 to Cys70 disulfide and substitutions for large effects on activity W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google Scholar). whereas substitutions that activity have effects on binding to and MMP-3, the of for Thr2 a in binding to to W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google Scholar). is in with the which indicate that Thr2 of TIMP-1 and of TIMP-2 with the region of the metalloproteinases that to the binding site for the residue of peptide the residue that has a dominant in MMP specificity K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar, C. W. J.J. A. H. K. J. PubMed Scopus Google Scholar). of a of the structural of TIMP-1 specificity and a generating variants that are more MMP inhibitors, have fifteen with substitutions for The results show that this residue has a influence on the specificity of TIMP for different metalloproteinases but show that is little between the of an amino 2 in TIMP-1 and the residue the site of a peptide substrate on activities with an MMP inhibitor substrate. The structural of these is results that protein of TIMP is a for generating more specific protein inhibitors of MMPs for of the roles of different MMPs and to the of for linked with excess activity of specific MMPs. The of targeted variants substitutions but the inhibitory of the and and TIMP-1 W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar, W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google indicate that residue 2 has a influence on specificity of the site The effects of substitutions for Thr2 about the of MMP binding by and TIMP variants with that the inhibitors for and MMP-3, on and have different substitutions The of for Thr2 a protein that is an inhibitor yet a native structure by Although the of TIMP not of the in TIMP-1 a protein that to the interactions of the C-terminal domain in a that has little for a active site and to the of TIMP is with cell and activity K. K. K. FEBS Lett. 1992; PubMed Scopus Google Scholar, K. A. J. Sci. 1994; PubMed Google Scholar, B. K. B. Murphy G. S. B. Sci. 1997; Google of the of residue 2 with the for different MMPs is to the structural of the effects of on the TIMP-1·MMP-3 the N-terminal region of has a different structure from that in K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). on the structure of that in TIMP during complex J. Freedman R.B. Williamson R.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). in the site the interaction with MMPs through effects on to different MMPs by from the disulfide The of for the that interactions of the S1′ pocket more of the binding for the interaction with with the structure of the S1′ pocket of is and those of MMPs. which in and MMP-3, into the S1′ pocket the in a more pocket Smith B. Biol. 1994; PubMed Scopus Google Scholar). The for the of for and threonine for example, and is a in the binding of to The specificity pocket of an MMP with substrate inhibitor to that the binding of TIMP by the ability of the residue 2 to with of in the of complexes S. Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar). has been that of the of each is on of the TIMP-1·MMP-3 complex K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). The C-terminal domain of TIMP-1 for of these and of the of interaction W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar). The results show that a residue of the site of a large influence on the and specificity of binding to MMPs. The of this is the of Thr2 from to on complex with the for H. Hooper N.M. Zinc Metalloproteinases in Health and Disease. Taylor & Francis Ltd., London1996: 153-204Google of the of the interaction site S. J. C. J. Biol. PubMed Scopus Google Scholar). The of of the of binding on of the Thr2 by the of that residue 2 of TIMP-1 a in the interaction a residue that has a large influence on the of the interaction 1995; 267: PubMed Scopus Google Scholar, Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar). form with in the structure K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google but of and and the of in that these are important for binding Thr2 W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google Scholar). the results show that the of the in the structure of the TIMP-1·MMP-3 complex are an important the structural of the inhibitory and specificity in but is important to to the stability of the The results that protein of TIMP is a for generating more specific protein inhibitors of MMPs for of the roles of different MMPs and to the of for linked with excess activity of specific MMPs. The of targeted variants substitutions but the inhibitory of the and and TIMP-1 W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar, W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google indicate that residue 2 has a influence on specificity of the site The effects of substitutions for Thr2 about the of MMP binding by and TIMP variants with that the inhibitors for and MMP-3, on and have different substitutions The of for Thr2 a protein that is an inhibitor yet a native structure by Although the of TIMP not of the in TIMP-1 a protein that to the interactions of the C-terminal domain in a that has little for a active site and to the of TIMP is with cell and activity K. K. K. FEBS Lett. 1992; PubMed Scopus Google Scholar, K. A. J. Sci. 1994; PubMed Google Scholar, B. K. B. Murphy G. S. B. Sci. 1997; Google Scholar). of the of residue 2 with the for different MMPs is to the structural of the effects of on the TIMP-1·MMP-3 the N-terminal region of has a different structure from that in K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). on the structure of that in TIMP during complex J. Freedman R.B. Williamson R.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). in the site the interaction with MMPs through effects on to different MMPs by from the disulfide The of for the that interactions of the S1′ pocket more of the binding for the interaction with with the MMPs. The structure of the S1′ pocket of is and those of MMPs. which in and MMP-3, into the S1′ pocket the in a more pocket Smith B. Biol. 1994; PubMed Scopus Google Scholar). The for the of for and threonine for example, and is a in the binding of to The specificity pocket of an MMP with substrate inhibitor to that the binding of TIMP by the ability of the residue 2 to with of in the of complexes S. Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar). has been that of the of each is on of the TIMP-1·MMP-3 complex K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google Scholar). The C-terminal domain of TIMP-1 for of these and of the of interaction W. Suzuki K. Nagase H. Arumugam S. Van Doren S.R. Brew K. FEBS Lett. 1996; 384: 155-161Crossref PubMed Scopus (97) Google Scholar). The results show that a residue of the site of a large influence on the and specificity of binding to MMPs. The of this is the of Thr2 from to on complex with the for H. Hooper N.M. Zinc Metalloproteinases in Health and Disease. Taylor & Francis Ltd., London1996: 153-204Google of the of the interaction site S. J. C. J. Biol. PubMed Scopus Google Scholar). The of of the of binding on of the Thr2 by the of that residue 2 of TIMP-1 a in the interaction a residue that has a large influence on the of the interaction 1995; 267: PubMed Scopus Google Scholar, Proc. Natl. Acad. Sci. U. S. A. 1996; PubMed Scopus Google Scholar). form with in the structure K. M. A. Suzuki K. N. Nagase H. Brew K. H. W. Nature. 1997; PubMed Scopus Google but of and and the of in that these are important for binding Thr2 W. Suzuki K. Nagase H. Brew K. J. Biol. Chem. 1997; 272: PubMed Scopus Google Scholar). the results show that the of the in the structure of the TIMP-1·MMP-3 complex are an important the structural of the inhibitory and specificity in but is important to to the stability of the of and of for to the of for that TIMP
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