Protein engineering and properties of human metalloproteinase and thrombospondin 1.
Protein engineering and properties of human metalloproteinase and thrombospondin 1.
复制标题
人金属蛋白酶和血小板反应蛋白的蛋白质工程和特性 1.
DOI：
10.1016/s0006-291x(02)00255-3
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发表时间：
2002
影响因子：
3.1
通讯作者：
Sang,Qing-XiangAmy
中科院分区：
文献类型：
--
作者：
Wei,Ping;Zhao,Yun-Ge;Zhuang,Li;Hurst,DouglasR;Ruben,Steve;Sang,Qing-XiangAmy
本工作产生了人金属蛋白酶和血小板反应蛋白1（METH-1或ADAMTS 1）的许多截短蛋白和Glu 385至Ala（E385/A）突变体以及特异性抗体。METH-1是一种活性内肽酶，金属蛋白酶和去整合素/富含半胱氨酸的结构域都是蛋白酶活性所必需的。在锌结合位点的点突变（E385/A）取消了催化活性。METH-1蛋白功能可通过多个位点的蛋白水解切割来调节。一个135 kDa的物种具有L33 GRPSEEDEE的NH 2-末端序列。在115 kDa处的一个种类和一些其他蛋白条带以F236 VSSHRYV 243开始，表明METH-1酶原可能被前蛋白转化酶如弗林蛋白酶通过切割R235-F236肽键而激活。这种切割不是自催化过程，因为E385/A突变体也被加工。此外，一个52 kDa的带L 800 KEPLTIQV的NH 2-末端序列之间的第一和第二血小板反应蛋白1样基序的胞外基质结合结构域的间隔区的消化。
This work generated many truncated proteins and Glu385to Ala (E385/A) mutants of the human metalloproteinase and thrombospondin 1 (METH-1 or ADAMTS1) and specific antibodies. METH-1 was an active endopeptidase and both the metalloproteinase and the disintegrin/cysteine-rich domains were required for the proteinase activity. A point mutation at the zinc-binding site (E385/A) abolished the catalytic activity. METH-1 protein function may be modulated through proteolytic cleavage at multiple sites. One 135 kDa species had an NH2-terminal sequence of L33GRPSEEDEE. A species at 115 kDa and some other protein bands began with F236VSSHRYV243, indicating that METH-1 proenzyme might be activated by a proprotein convertase such as furin by cleaving the R235–F236peptide bond. This cleavage was not an autocatalytic process since the E385/A mutants were also processed. Furthermore, a 52 kDa band with an NH2-terminal sequence of L800KEPLTIQV resulted from the digestion between the first and the second thrombospondin 1-like motifs in the spacer region of the extracellular matrix-binding domains.