Osthues A, Knäuper V, Oberhoff R, Reinke H, Tschesche H
University of Bielefeld, Faculty of Chemistry, Department of Biochemistry, Germany.
FEBS Lett. 1992 Jan 13;296(1):16-20. doi: 10.1016/0014-5793(92)80393-u.
The tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2 were purified to apparent homogeneity from human rheumatoid synovial fluid (HRSF). The inhibitors were isolated by dissociation of non-covalent gelatinase/TIMP complexes. TIMP-1 migrated as a single polypeptide with Mr 28,500 on SDS-PAGE, while the Mr of TIMP-2 was 21,000. The inhibitory activity was stable under heat and acid pH. N-terminal sequence data were obtained for the first 15 residues of both inhibitors and showed identity to the human fibroblast inhibitors TIMP-1 and TIMP-2. This is the first demonstration that TIMP-1 and TIMP-2 can be directly purified from human rheumatoid synovial fluid. The complex formation between the metalloproteinase inhibitors and leucocyte metalloproteinases was shown by immunoblotting.
金属蛋白酶组织抑制剂TIMP-1和TIMP-2从人类风湿性滑液（HRSF）中纯化至表观均一。通过解离非共价明胶酶/TIMP复合物来分离这些抑制剂。TIMP-1在SDS-PAGE上以单一分子量为28,500的多肽形式迁移，而TIMP-2的分子量为21,000。其抑制活性在加热和酸性pH条件下稳定。获得了两种抑制剂前15个残基的N端序列数据，显示与人成纤维细胞抑制剂TIMP-1和TIMP-2相同。这是首次证明TIMP-1和TIMP-2可直接从人类风湿性滑液中纯化。免疫印迹显示了金属蛋白酶抑制剂与白细胞金属蛋白酶之间的复合物形成情况。
