Regulation of the α-secretase ADAM10 by its prodomain and proprotein convertases
Regulation of the α-secretase ADAM10 by its prodomain and proprotein convertases
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DOI：
10.1096/fj.01-0007fje
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发表时间：
2001-06-01
期刊：
影响因子：
4.8
通讯作者：
Fahrenholz, F
中科院分区：
文献类型：
--
作者：
Anders, A;Gilbert, S;Fahrenholz, F
阿尔茨海默病淀粉样前体蛋白的胞外域脱落由α-和β -分泌酶介导，而这两种酶自身也受到蛋白水解加工。解整合素金属蛋白酶ADAM10作为一种酶原合成，在其前结构域和催化结构域之间具有一个前蛋白转化酶（PC）识别序列。在本研究中，我们探讨了前结构域在调节ADAM10的α -分泌酶活性中的作用。人前蛋白转化酶PC7和弗林蛋白酶在人胚胎肾293细胞中的过表达显示ADAM10成熟增加，导致淀粉样前体蛋白的α -分泌酶介导的加工增强。ADAM10中PC识别序列的突变以及使用PC抑制剂和弗林蛋白酶缺陷的LoVo细胞系证实了PC，特别是PC7在ADAM10成熟和激活中的作用。此外，我们证明了ADAM10的前结构域具有双重功能。当作为单独的多肽反式共表达时，它抑制野生型ADAM10的α -分泌酶活性。然而，前结构域作为一种分子伴侣，功能性地挽救了一种缺失前结构域的先前无活性的ADAM10突变体的α -分泌酶活性。我们的研究结果为增强非淀粉样生成的α -分泌酶途径提出了新的方法。
Ectodomain shedding of the Alzheimer's amyloid precursor protein is mediated by alpha- and beta -secretases, which, for their part, are also proteolytically processed. The disintegrin metalloproteinase ADAM10 is synthesized as a zymogen with a proprotein convertase (PC) recognition sequence between the prodomain and the catalytic domain. In this study, we investigated the role of the prodomain in the regulation of the alpha -secretase activity of ADAM10. Overexpression of the proprotein convertases PC7 and furin in human embryonic kidney 293 cells revealed an increased ADAM10 maturation resulting in enhanced alpha -secretase-mediated processing of amyloid precursor protein. Mutation of the PC recognition sequence in ADAM10 as well as the use of a PC inhibitor and of the furin-deficient LoVo cell line confirmed the role of PCs, in particular, of PC7, in ADAM10 maturation and activation. Furthermore, we demonstrated that the prodomain of ADAM10 has a dual function. When coexpressed in trans as separate polypeptide, it inhibited the alpha -secretase activity of wild-type ADAM10. However, the prodomain acted as a chaperone and functionally rescued the alpha -secretase activity of a former inactive ADAM10 mutant lacking the prodomain. The results of our study suggest new approaches to enhance the nonamyloidogenic alpha -secretase pathway.