应用生物功能化色谱研究SSAO酶与底物和抑制剂之间的相互作用

Investigation on the Interaction Between SSAO,Its Substrate and the Inhibitor Using Bio-Functionalized Chromatography

应用生物功能化色谱技术——毛细管电泳法研究对氨基脲敏感的胺氧化酶（SSAO）与其底物苯甲胺、抑制剂2-溴乙胺之间的相互作用．活性检测表明，SSAO酶经固定化到脂质体上之后，仍然保留有70％～85％的活性．将有活性的固定化酶添加到磷酸盐缓冲液中，随着缓冲液中固定化酶浓度的增加，pH=5时，特异性底物苯甲胺的有效淌度从4．06×10^-4cm^2·V^-1·s^-1下降到0．81×10^-4cm^2·V^-1·S^-1，pH=7时，则从3．91×10^-4cm^2·V^-1·S^-1下降到0．29×10^-4cm^2·V^-1·s^-1．若将抑制剂2-溴乙胺与固定化酶同时添加到电泳缓冲液中，随着抑制剂浓度从10^-6mol·L^-1上升到10^-1mol·L^-1，苯甲胺的有效淌度则从1．42×10^-4cm^2·V^-1·s^-1上升到2．22×10^-4cm·V^-2·s^-1．

Bio-functionalized chromatography--capillary electrophoresis was used in investigating the interaction between semiearbazide-sensitive amine oxidase（SSAO）, its substrate benzylamine as well as its inhibitor 2-BrEA. SSAO activity detection showed that SSAO could keep 70% 85% activity of its free form after being immobilized onto liposome. The active immobilized SSAO was added to the PBS. With the increase of the immobilized enzyme concentration, the effective mobility of its specific substrate, benzylamine, decreased from 4.06 × 10^-4 cm^2 · V^-1· s^-1 to 0. 81×10^-4cm^2· V^-1·s^-1 at pH=5, and from 3.91×10^-4cm^2·V^-1·s^-1 to 0. 29×10^-4cm^2 · V^-1· s^-1 at pH=7. If the concentration of the inhibitor 2-BrEA in the buffer was increased from 10^-6mol · L^-1 to 10^-1mol · L^-1 , the effective mobility of benzylamine would increase from 1. 42×10^-4cm^-2· V^-1·s^-1 to 2.22×10^-4cm^2 · V^-1· s^-1.