Human tissue inhibitor of metalloproteinases 3 interacts with both the N- and C-terminal domains of gelatinases A and B - Regulation by polyanions
Human tissue inhibitor of metalloproteinases 3 interacts with both the N- and C-terminal domains of gelatinases A and B - Regulation by polyanions
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DOI：
10.1074/jbc.274.16.10846
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发表时间：
1999-04-16
影响因子：
4.8
通讯作者：
Murphy, G
中科院分区：
文献类型：
--
作者：
Butler, GS;Apte, SS;Murphy, G
我们使用连续测定将金属蛋白酶组织抑制剂 (TIMP)-3 与各种基质金属蛋白酶的关联常数与 TIMP-1 和 TIMP-2 的关联常数进行了比较。 TIMP-3 的行为比 TIMP-1 更像 TIMP-3，显示出与明胶酶 A 和 B 的快速关联。使用明胶酶 A 的 N 末端结构域、分离的 C 末端结构域或无活性的原明胶酶 A 突变体进行的实验表明，明胶酶 A 的血红素结合蛋白结构域对与 TIMP-3 的相互作用做出了重要贡献。明胶酶 A 血红素结合蛋白结构域的部分与溶基质素结构域的部分交换表明，明胶酶 A 的残基 568-631 是与 TIMP-3 快速结合所必需的。单独的明胶酶 B 的 N 端结构域也显示出与 TIMP-3 的较慢关联，再次暗示显着的 C 结构域相互作用。在明胶-琼脂糖上分离 TIMP-3 与原明胶酶 A 和 B 之间的复合物证明 TIMP-3 与两种酶原结合。我们在可溶性测定中分析了各种聚阴离子对 TIMP-3 抑制活性的影响。硫酸葡聚糖、肝素和硫酸乙酰肝素可增加关联率，但硫酸皮肤素或透明质酸则不会增加关联率。由于 TIMP-3 被隔离在细胞外基质中，某些硫酸乙酰肝素蛋白聚糖的存在可以增强其抑制能力。
We compared the association constants of tissue inhibitor of metalloproteinases (TIMP)-3 with various matrix metalloproteinases with those for TIMP-1 and TIMP-2 using a continuous assay. TIMP-3 behaved more like TIMP-3 than TIMP-1, showing rapid association with gelatinases A and B. Experiments with the N-terminal domain of gelatinase A the isolated C-terminal domain, or an inactive progelatinase A mutant showed that the hemopexin domain of gelatinase A makes an important contribution to the interaction with TIMP-3. The exchange of portions of the gelatinase A hemopexin domain with that of stromelysin revealed that residues 568-631 of gelatinase A were required for rapid association with TIMP-3. The N-terminal domain of gelatinase B alone also showed slower association with TIMP-3, again implying significant C-domain interactions. The isolation of complexes between TIMP-3 and progelatinases A and B on gelatin-agarose demonstrated that TIMP-3 binds to both proenzymes. We analyzed the effect of various polyanions on the inhibitory activity of TIMP-3 in our soluble assay. The association rate was increased by dextran sulfate, heparin, and heparan sulfate, but not by dermatan sulfate or hyaluronic acid. Because TIMP-3 is sequestered in the extracellular matrix, the presence of certain heparan sulfate proteoglycans could enhance its inhibitory capacity.