Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA.
J Am Chem Soc. 2013 Jun 19;135(24):9139-48. doi: 10.1021/ja403523p. Epub 2013 Jun 6.
Matrix metalloproteinases (MMPs) are zinc endopeptidases that play roles in numerous pathophysiological processes and therefore are promising drug targets. However, the large size of this family and a lack of highly selective compounds that can be used for imaging or inhibition of specific MMPs members has limited efforts to better define their biological function. Here we describe a protein engineering strategy coupled with small-molecule probe design to selectively target individual members of the MMP family. Specifically, we introduce a cysteine residue near the active-site of a selected protease that does not alter its overall activity or function but allows direct covalent modification by a small-molecule probe containing a reactive electrophile. This specific engineered interaction between the probe and the target protease provides a means to both image and inhibit the modified protease with absolute specificity. Here we demonstrate the feasibility of the approach for two distinct MMP proteases, MMP-12 and MT1-MMP (or MMP-14).
基质金属蛋白酶（MMPs）是锌内肽酶，在许多病理生理过程中发挥作用，因此是有前途的药物靶点。然而，由于这个家族的规模庞大，以及缺乏可用于成像或抑制特定 MMP 成员的高度选择性化合物，限制了人们对更好地定义其生物学功能的努力。在这里，我们描述了一种蛋白质工程策略，结合小分子探针设计，以选择性地靶向 MMP 家族的个别成员。具体来说，我们在选定的蛋白酶的活性位点附近引入一个半胱氨酸残基，该残基不会改变其整体活性或功能，但允许含有反应性亲电试剂的小分子探针直接共价修饰。探针与靶蛋白酶之间的这种特定的工程化相互作用为用绝对特异性对修饰的蛋白酶进行成像和抑制提供了一种手段。在这里，我们证明了该方法对于两种不同的 MMP 蛋白酶 MMP-12 和 MT1-MMP（或 MMP-14）的可行性。
