Structural Basis for Matrix Metalloproteinase-2 (MMP-2)-selective Inhibitory Action of β-Amyloid Precursor Protein-derived Inhibitor
Structural Basis for Matrix Metalloproteinase-2 (MMP-2)-selective Inhibitory Action of β-Amyloid Precursor Protein-derived Inhibitor
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DOI：
10.1074/jbc.m111.264176
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发表时间：
2011-09-23
影响因子：
4.8
通讯作者：
Higashi, Shouichi
中科院分区：
文献类型：
--
作者：
Hashimoto, Hiroshi;Takeuchi, Tomoka;Higashi, Shouichi
与其他合成或生理抑制基质金属蛋白酶(MMPs)的药物不同，具有ISYGNDALMP序列的β-淀粉样前体蛋白衍生抑制肽(APP-IP)对MMP2具有很高的选择性。我们先前的研究确定了APP-IP选择性抑制MMP2所必需的氨基酸残基，并证明了十肽抑制剂相对于底物结合裂解的N到C方向与底物相反。然而，这两个分子之间的详细相互作用仍有待澄清。在此，我们测定了与APP-IP形成的复合体中基质金属蛋白酶-2的催化域的晶体结构。我们发现，复合体中的APP-IP确实嵌入了催化结构域的底物结合裂隙中，方向与底物相反。结合晶体结构，首次明确了该抑制剂Tyr(3)的芳香族侧链位于该酶的S1‘口袋中，APP-IP的天冬氨酸(6)的羧酸基与该酶的催化锌发生双向配位。抑制剂的Ala(7)-Pro(10)和Tyr(3)-Ile(1)链分别延伸到裂隙的非质侧和质侧。因此，十肽抑制剂与酶的底物结合裂隙有很长的接触范围。这种相互作用模式可能是抑制物高选择性的关键，因为MMPs在催化中心附近具有共同的结构，但其底物结合裂隙的整个结构具有足够的多样性，使抑制剂能够将MMP2与其他MMPs区分开来。
Unlike other synthetic or physiological inhibitors for matrix metalloproteinases (MMPs), the beta-amyloid precursor protein-derived inhibitory peptide (APP-IP) having an ISYGNDALMP sequence has a high selectivity toward MMP-2. Our previous study identified amino acid residues of MMP-2 essential for its selective inhibition by APP-IP and demonstrated that the N to C direction of the decapeptide inhibitor relative to the substrate-binding cleft of MMP-2 is opposite that of substrate. However, detailed interactions between the two molecules remained to be clarified. Here, we determined the crystal structure of the catalytic domain of MMP-2 in complex with APP-IP. We found that APP-IP in the complex is indeed embedded into the substrate-binding cleft of the catalytic domain in the N to C direction opposite that of substrate. With the crystal structure, it was first clarified that the aromatic side chain of Tyr(3) of the inhibitor is accommodated into the S1' pocket of the protease, and the carboxylate group of Asp(6) of APP-IP coordinates bidentately to the catalytic zinc of the enzyme. The Ala(7) to Pro(10) and Tyr(3) to Ile(1) strands of the inhibitor extend into the nonprime and the prime sides of the cleft, respectively. Therefore, the decapeptide inhibitor has long range contact with the substrate-binding cleft of the protease. This mode of interaction is probably essential for the high MMP-2 selectivity of the inhibitor because MMPs share a common architecture in the vicinity of the catalytic center, but whole structures of their substrate-binding clefts have sufficient variety for the inhibitor to distinguish MMP-2 from other MMPs.