酶动力学_____蛋白激酶CK2；_重组蛋白；

藤黄菌素体外对重组人蛋白激酶CK2全酶的抑制作用

http://www.100md.com 《第四军医大学学报》 2006年第7期

酶动力学,,蛋白激酶CK2；,重组蛋白；,全酶；,藤黄菌素；,酶动力学,0引言,1材料和方法,2结果,3讨论,【参考文献】

     Inhibitory effect of luteolin on recombinant human protein kinase CK2 holoenzyme in vitro

    LIN XiaoCong, LIU XinGuang, CHEN XiaoWen, CHEN WeiZhu, LIANG NianCi

    Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang 524023, China, Institute of Pediatrics, Shenzhen Childrens Hospital, Shenzhen 518026, China

    【Abstract】 AIM: To observe the inhibitory effect of luteolin on recombinant human protein kinase CK2 holoenzyme and to confirm the inhibitory type by its enzyme kinetic analysis in vitro. METHODS: Recombinant human protein kinase CK2 α′ and β subunits were cloned, expressed and purified. The 2 subunits were mixed at equal molar ratio to reconstitute CK2 holoenzyme. Luteolin inhibitory effect was detected by determining the radioactivity of 32P of [γ32P] ATP incorporated into the CK2 substrate and CK2 kinetic analysis was carried out by using the LineweaverBurk plot. RESULTS: Luteolin was proved to inhibit strongly the holoenzyme activity of recombinant human protein kinase CK2 with IC50 of 0.86 μmol/L. Kinetic studies of luteolin on recombinant human CK2 showed that luteolin acted as an inhibitor, competitive with ATP (Ki=0.19 μmol/L) and cooperative with casein (Ki=0.11 μmol/L). CONCLUSION: Luteolin is an effective inhibitor of protein kinase CK2 in vitro. ......

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