Structural determinants of the ADAM inhibition by TIMP-3: crystal structure of the TACE-N-TIMP-3 complex |
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Authors: | Wisniewska Magdalena Goettig Peter Maskos Klaus Belouski Edward Winters Dwight Hecht Randy Black Roy Bode Wolfram |
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Institution: | 1 Max-Planck-Institut für Biochemie, Proteinase Research Group, Am Klopferspitz 18, D-82152 Martinsried, Germany 2 Present address: Proteros Biostructures GmbH, Am Klopferspitz 19, D-82152 Martinsried, Germany 3 Amgen, Department of Protein Sciences,Thousand Oaks, CA, USA 4 Amgen, Department of Inflammation, 1201 Amgen Court West, Seattle, WA 98119-3105, USA |
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Abstract: | TIMP-3 (tissue inhibitor of metalloproteinases 3) is unique among the TIMP inhibitors, in that it effectively inhibits the TNF-α converting enzyme (TACE). In order to understand this selective capability of inhibition, we crystallized the complex formed by the catalytic domain of recombinant human TACE and the N-terminal domain of TIMP-3 (N-TIMP-3), and determined its molecular structure with X-ray data to 2.3 Å resolution. The structure reveals that TIMP-3 exhibits a fold similar to those of TIMP-1 and TIMP-2, and interacts through its functional binding edge, which consists of the N-terminal segment and other loops, with the active-site cleft of TACE in a manner similar to that of matrix metalloproteinases (MMPs). Therefore, the mechanism of TIMP-3 binding toward TACE is not fundamentally different from that previously elucidated for the MMPs. The Phe34 phenyl side chain situated at the tip of the relatively short sA-sB loop of TIMP-3 extends into a unique hydrophobic groove of the TACE surface, and two Leu residues in the adjacent sC-connector and sE-sF loops are tightly packed in the interface allowing favourable interactions, in agreement with predictions obtained by systematic mutations by Gillian Murphy's group. The combination of favourable functional epitopes together with a considerable flexibility renders TIMP-3 an efficient TACE inhibitor. This structure might provide means to design more efficient TIMP inhibitors of TACE. |
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Keywords: | ADAM a disintegrin and metalloproteinase ADAMTS ADAMs with thrombospondin motifs β-APP amyloid precursor protein cd catalytic domain ECM extracellular matrix MMP matrix metalloproteinase TACE TNF-α converting enzyme TIMP tissue inhibitor of metalloproteinases N-TIMP N-terminal domain of TIMP TNF-α tumor necrosis factor-α VAP vascular apoptosis-inducing protein |
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