Cell surface binding of TIMP‐2 and pro‐MMP‐2/TIMP‐2 complex

FEBS Letters

Corcoran, Marta L.¹  Foidart, Jean-Michel²  Krutzsch, Henry C.¹  Emonard, HervéP.²  Stetler-Stevenson, William G.¹  Emmert-Buck, Michael R.¹ 

[1] Laboratory of Pathology, Extracellular Matrix Pathology Section, National Cancer Institute, Bethesda, MD 20892, USA;Laboratory of Biology, University of Liège, Liège, Belgium

关键词: Metalloproteinase;    Tumor cell;    Invasion;    Activation;    Tissue Inhibitor;    TIMP-2;    MMP;    matrix metalloproteinase;    TIMP;    tissue inhibitor of metalloproteinase;    rTIMP-2;    recombinant TIMP-2;    PBS;    phosphatebuffered saline;    BSA;    bovine serum albumin;    HEPES;    4-(2-hydroxyethyl)-1-piperazinethanesulfonic acid;    SDS;    sodium dodecyl sulfate;    DMEM;    Dulbecc'os modified eagles medium;    FBS;    fetal bovine serum;    dpm;    disintegrations per minute;   

DOI  :  10.1016/0014-5793(95)00345-A

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Tissue inhibitor of metalloproteinases (TIMP-2) is a low molecular weight proteinase inhibitor capable of inhibiting activated matrix metalloproteinases (MMPs). TIMP-2 is found both free and in a 1:1 stoichiometric complex with the pro-enzyme form of MMP-2 (pro-MMP-2/TIMP-2 complex). We have measured the binding of recombinant TIMP-2 to intact HT-1080 and MCF-7 cells. HT-1080 cells in suspension bound ¹²⁵I-labeled rTIMP-2 with a K _d of 2.5 nM and 30,000 sites/cell. Monolayers of MCF-7 cells were similarly found to bind [¹²⁵I]rTIMP-2 with a K _d of 1.6 nM and 25,000 sites/cell. Specific binding of MMP-2 alone to HT-1080 cells was not observed; however, pro-MMP-2/TIMP-2 complex was capable of binding to the surface of HT-1080 cells in a TIMP-2-dependent manner. Binding of rTIMP-2 was not competed by the presence of TIMP-1. These results suggest that rTIMP-2 alone binds directly to the cell surface of HT-1080 and MCF-7 cell lines, and TIMP-2 is capable of localizing MMP-2 to the surface of HT-1080 cells via interaction with a specific binding site.

【 授权许可】

Unknown