Biophysical and Functional Characterization of Full-length, Recombinant Human Tissue Inhibitor of Metalloproteinases-2 (TIMP-2) Produced in Escherichia coli COMPARISON OF WILD TYPE AND AMINO-TERMINAL ALANINE APPENDED VARIANT WITH IMPLICATIONS FOR THE MECHA

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作者：

PT Wingfield，JK Sax，SJ Stahl，J Kaufman，I Palmer，V Chung，ML Corcoran，DE Kleiner，WG Stetlerstevenson

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摘要：

Matrix metalloproteinases (MMPs) function in the remodeling of the that is integral for many normal and pathological processes. The family, including (), regulates the activity of these multifunctional metalloproteinases. family members are that contain six conserved bonds, one involving an amino-terminal that is critical for MMP inhibitor activity. has been expressed in , folded from insoluble protein, and functionally characterized. The wild type protein inhibited (), whereas a variant with an alanine appended to the amino terminus (Ala+) was inactive. Removal of amino-terminal alanine by exopeptidase restored activity. This confirms the mechanistic importance of the amino-terminal amino group in the metalloproteinase inhibitory activity, as originally suggested from the x-ray structure of a complex of with and a complex of with -MMP. The Ala+variant exhibited conformational, pro-complex formation and fibroblast growth modulating properties of the wild type protein. These findings demonstrate that Ala+is an excellent biochemical tool for examining the specific role of MMP inhibition in the multiple functions ascribed to .

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关键词：

Environmental law

DOI：

10.1074/jbc.274.30.21362

被引量：

247

年份：

1999