Murphy G, Houbrechts A, Cockett M I, Williamson R A, O'Shea M, Docherty A J
Strangeways Research Laboratory, Cambridge, U.K.
Biochemistry. 1991 Aug 20;30(33):8097-102. doi: 10.1021/bi00247a001.
Recombinant tissue inhibitor of metalloproteinases (TIMP-1) and a truncated version containing only the three N-terminal loops, delta 127-184TIMP, have been expressed in myeloma cells and purified by affinity chromatography and gel filtration. delta 127-184TIMP was found to exist as two main glycosylation variants of molecular mass 24 kD and 19.5 kDa and an unglycosylated form of 13 kDa. All forms of the truncated inhibitor were able to inhibit and form complexes with active forms of the matrix metalloproteinases, indicating that the major structural features for specific interaction with these enzymes resides in these three loops. Stable binding of delta 127-184TIMP to pro 95-kDa gelatinase was not demonstrable under the conditions for binding of full-length TIMP-1.
重组金属蛋白酶组织抑制剂（TIMP-1）以及仅包含三个N端环的截短版本delta 127-184TIMP已在骨髓瘤细胞中表达，并通过亲和色谱和凝胶过滤进行纯化。发现delta 127-184TIMP以分子量分别为24 kD和19.5 kDa的两种主要糖基化变体以及13 kDa的未糖基化形式存在。截短抑制剂的所有形式都能够与基质金属蛋白酶的活性形式结合并形成复合物，这表明与这些酶特异性相互作用的主要结构特征存在于这三个环中。在全长TIMP-1结合的条件下，未证实delta 127-184TIMP与95 kDa前明胶酶的稳定结合。
