Rosendahl M S, Ko S C, Long D L, Brewer M T, Rosenzweig B, Hedl E, Anderson L, Pyle S M, Moreland J, Meyers M A, Kohno T, Lyons D, Lichenstein H S
Amgen Inc., Boulder, Colorado 80301, USA.
J Biol Chem. 1997 Sep 26;272(39):24588-93. doi: 10.1074/jbc.272.39.24588.
Tumor necrosis factor-alpha (TNF) is initially expressed as a 26-kDa membrane-bound precusor protein (pro-TNF) that is shed proteolytically from the cell surface, releasing soluble 17-kDa TNF. We have identified human ADAM 10 (HuAD10) from THP-1 membrane extracts as a metalloprotease that specifically clips a peptide substrate spanning the authentic cleavage site between Ala76 and Val77 in pro-TNF. To confirm that HuAD10 has TNF processing activity, we cloned, expressed, and purified an active, truncated form of HuAD10. Characterization of recombinant HuAD10 (rHuAD10) suggests that this enzyme has many of the properties (i.e. substrate specificity, metalloprotease activity, cellular location) expected for a physiologically relevant TNF-processing enzyme.
肿瘤坏死因子-α（TNF）最初表达为一种26 kDa的膜结合前体蛋白（pro-TNF），该蛋白从细胞表面被蛋白水解切割，释放出可溶性的17 kDa TNF。我们从THP-1膜提取物中鉴定出人类ADAM 10（HuAD10）作为一种金属蛋白酶，它能特异性切割跨越pro-TNF中Ala76和Val77之间真实切割位点的肽底物。为了证实HuAD10具有TNF加工活性，我们克隆、表达并纯化了一种活性截短形式的HuAD10。重组HuAD10（rHuAD10）的特性表明，这种酶具有作为生理相关TNF加工酶所预期的许多特性（即底物特异性、金属蛋白酶活性、细胞定位）。
