Kinetic analysis of the binding of hemopexin-like domain of gelatinase B cloned and expressed in Pichia pastoris to tissue inhibitor of metalloproteinases-1.
Kinetic analysis of the binding of hemopexin-like domain of gelatinase B cloned and expressed in Pichia pastoris to tissue inhibitor of metalloproteinases-1.
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毕赤酵母中克隆和表达的明胶酶 B 的血红素结合蛋白样结构域与金属蛋白酶-1 组织抑制剂结合的动力学分析。
DOI：
10.1023/b:jopc.0000005499.51466.50
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Tschesche,Harald
中科院分区：
文献类型：
--
作者：
Stute,Jörg;Pourmotabbed,Tayebeh;Tschesche,Harald
明胶酶是基质金属蛋白酶家族的一个亚类。它们的C末端类血凝蛋白结构域与金属蛋白酶组织抑制因子(TIMP)的相互作用是明胶酶调控机制的主要部分。为了研究明胶酶B(92-PEX)的血凝蛋白结构域与TIMP-1的相互作用，我们在巴斯德毕赤酵母中表达了该结构域。活性复性结构域通过离子交换层析和凝胶过滤得到纯化。我们用表面等离子体共振(SPR)研究了92-Pex/TIMP-1复合体的形成。解离常数Kd值为0.86 nm。与明胶酶A的血粘蛋白样结构域与TIMP-2的复合物(Olson，M.W.等人，1997)类似，92-Pex/TIMP-1复合物的结合曲线最好地符合单相模型。
The gelatinases are a subgroup of the matrix metalloproteinase family. The interaction of their C-terminal hemopexin-like domain with a tissue inhibitor of metalloproteinases (TIMP) is a major part of the regulatory mechanisms of gelatinases. To investigate the interaction of the hemopexinlike domain of gelatinase B (92-Pex) and TIMP-1, we expressed the individual domain inPichia pastoris. The active refolded domain was purified by ion exchange chromatography and gel filtration. We investigated the formation of the 92-Pex/TIMP-1 complex by surface plasmon resonance (SPR). The dissociation constant Kdwas calculated to be 0.86 nM. Analogous to the complex of the hemopexin-like domain of gelatinase A and TIMP-2 (Olson, M. W.et al., 1997), the binding curves of the 92-Pex/TIMP-1 complex were best fitted with a monophasic model.