Kinetic Analysis of the Binding of Hemopexin-Like Domain of Gelatinase B Cloned and Expressed in Pichia pastoris to Tissue Inhibitor of Metalloproteinases-1.

Autor: Stute, Jörg, Pourmatabbed, Tayebeh, Tschesche, Harald
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Zdroj: Journal of Protein Chemistry; Aug2003, Vol. 22 Issue 6, p509-514, 6p
Abstrakt: 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 hemopexin-like domain of gelatinase B (92-Pex) and TIMP-1, we expressed the individual domain in Pichia 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 Kd was 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. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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