Matrix metalloproteinase-10 (MMP-10) interaction with tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2: Binding studies and crystal structure

Department of Cancer Biology, Mayo Clinic Comprehensive Cancer Center, Jacksonville, Florida 32224, USA.
Journal of Biological Chemistry (Impact Factor: 4.57). 03/2012; 287(19):15935-46. DOI: 10.1074/jbc.M112.341156
Source: PubMed


Matrix metalloproteinase 10 (MMP-10, stromelysin-2) is a secreted metalloproteinase with functions in skeletal development, wound healing, and vascular remodeling; its overexpression is also implicated in lung tumorigenesis and tumor progression. To understand the regulation of MMP-10 by tissue inhibitors of metalloproteinases (TIMPs), we have assessed equilibrium inhibition constants (K(i)) of putative physiological inhibitors TIMP-1 and TIMP-2 for the active catalytic domain of human MMP-10 (MMP-10cd) using multiple kinetic approaches. We find that TIMP-1 inhibits the MMP-10cd with a K(i) of 1.1 × 10(-9) M; this interaction is 10-fold weaker than the inhibition of the similar MMP-3 (stromelysin-1) catalytic domain (MMP-3cd) by TIMP-1. TIMP-2 inhibits the MMP-10cd with a K(i) of 5.8 × 10(-9) M, which is again 10-fold weaker than the inhibition of MMP-3cd by this inhibitor (K(i) = 5.5 × 10(-10) M). We solved the x-ray crystal structure of TIMP-1 bound to the MMP-10cd at 1.9 Å resolution; the structure was solved by molecular replacement and refined with an R-factor of 0.215 (R(free) = 0.266). Comparing our structure of MMP-10cd·TIMP-1 with the previously solved structure of MMP-3cd·TIMP-1 (Protein Data Bank entry 1UEA), we see substantial differences at the binding interface that provide insight into the differential binding of stromelysin family members to TIMP-1. This structural information may ultimately assist in the design of more selective TIMP-based inhibitors tailored for specificity toward individual members of the stromelysin family, with potential therapeutic applications.

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    • "Of particular interest were the genes encoding matrix metalloproteinase 10 (MMP10), also referred to as stromelysin 2, which was upregulated by 3.3-fold, and its inhibitor tissue inhibitor of metalloproteinase 2 (TIMP2), which was downregulated by 3.2-fold. MMP10 is a secreted proteinase that degrades the extracellular matrix, and TIMP2 inhibits MMP10 activity [22]. Thus, the upregulation of MMP10 and concomitant downregulation of TIMP2 would be predicted to disrupt cell-cell adhesion and promote invasion and metastasis of YY1 knockdown BXPC-3 cells. "
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    Molecular Cancer 05/2014; 13(1):130. DOI:10.1186/1476-4598-13-130 · 4.26 Impact Factor
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    • "Mature secreted full-length rhTIMP-1 was expressed using the pTT/TIMP-1 construct transfected into HEK 293E cells [30], and was purified by SP-Sepharose chromatography as we have described previously [31]. Four mutant pTT/TIMP-1 constructs were generated to introduce a free Cys residue (R180C, S181C, Q182C, and A184C) using the Stratagene QuikChange mutagenesis kit (Agilent Technologies, Wilmington, DE, USA) according to the manufacturer’s protocols. "
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