Moss Marcia L, Bomar Martha, Liu Qian, Sage Harvey, Dempsey Peter, Lenhart Patricia M, Gillispie Patricia A, Stoeck Alexander, Wildeboer Dirk, Bartsch Jörg W, Palmisano Ralf, Zhou Pei
BioZyme Incorporated, Apex, North Carolina 27523, USA.
J Biol Chem. 2007 Dec 7;282(49):35712-21. doi: 10.1074/jbc.M703231200. Epub 2007 Sep 25.
ADAM10 is a disintegrin metalloproteinase that processes amyloid precursor protein and ErbB ligands and is involved in the shedding of many type I and type II single membrane-spanning proteins. Like tumor necrosis factor-alpha-converting enzyme (TACE or ADAM17), ADAM10 is expressed as a zymogen, and removal of the prodomain results in its activation. Here we report that the recombinant mouse ADAM10 prodomain, purified from Escherichia coli, is a potent competitive inhibitor of the human ADAM10 catalytic/disintegrin domain, with a K(i) of 48 nM. Moreover, the mouse ADAM10 prodomain is a selective inhibitor as it only weakly inhibits other ADAM family proteinases in the micromolar range and does not inhibit members of the matrix metalloproteinase family under similar conditions. Mouse prodomains of TACE and ADAM8 do not inhibit their respective enzymes, indicating that ADAM10 inhibition by its prodomain is unique. In cell-based assays we show that the ADAM10 prodomain inhibits betacellulin shedding, demonstrating that it could be of potential use as a therapeutic agent to treat cancer.
ADAM10是一种去整合素金属蛋白酶，可加工淀粉样前体蛋白和表皮生长因子受体（ErbB）配体，并参与多种I型和II型单跨膜蛋白的脱落。与肿瘤坏死因子-α转化酶（TACE或ADAM17）一样，ADAM10以酶原形式表达，前结构域的去除会导致其激活。在此我们报告，从大肠杆菌中纯化的重组小鼠ADAM10前结构域是人类ADAM10催化/去整合素结构域的有效竞争性抑制剂，抑制常数（K(i)）为48 nM。此外，小鼠ADAM10前结构域是一种选择性抑制剂，因为它仅在微摩尔范围内微弱抑制其他ADAM家族蛋白酶，且在类似条件下不抑制基质金属蛋白酶家族成员。TACE和ADAM8的小鼠前结构域不抑制它们各自的酶，这表明ADAM10前结构域对其的抑制作用是独特的。在基于细胞的实验中，我们表明ADAM10前结构域抑制β细胞ulin的脱落，证明它可能作为治疗癌症的治疗剂具有潜在用途。
