Overview:
CDK1 or Cell Division Control protein 1 is essential for the completion of START, the controlling event in the cell cycle that is required to initiate mitosis. CDK1 is a catalytic subunit of a protein kinase complex, called the M-Phase Promoting Factor that induces entry into mitosis and is universal among eukaryotes (1). Phosphorylation of Bcl-2 in G2/M phase-arrested cells following photodynamic therapy with hypericin involves a CDK1-mediated signal and delays the onset of apoptosis. Therapeutic potential of CDK inhibitor NU2058 in androgen-independent prostate cancer has also been demonstrated (2).
Gene Aliases:
CDK1: CDC2 CyclinB1: CCNB1, CCNB
Genbank Number:
NM_001786NM_031966
References:
1. Vantieghem, A. et al: Phosphorylation of Bcl-2 in G2/M phase-arrested cells following photodynamic therapy with hypericin involves a CDK1-mediated signal and delays the onset of apoptosis. J. Biol. Chem. 2002; 277(40):37718-31.2. Rigas, A.C. et al: Therapeutic potential of CDK inhibitor NU2058 in androgen-independent prostate cancer. Oncogene. 2007; 18.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
1.对于抑制剂筛选工作(求ic50)是不是体系内酶与底物的量(底物应该是过量的)对实验结果影响不大。
2.如果要求算Km值,是不是需要知道反应产物的绝对量。反应时间文献上都是5分钟,反应速度就用反应产物量除以反应时间即可。
3.酶是进口分装的,规格5U,一次用不完,用PBS稀释后如何保存
谢谢
1、测定酶比活力:底物需要过量么?测定时间多长?是否可以加入过量的底物,然后测定3min吸光度的增加值,从吸光度的变化值计算比活力。
2、在酶抑制剂筛选的过程中,是否需要保证底物过量?还是要水浴一定时间让反应完全?我看到文献说用终浓度为0.1μmol/mL的底物,终浓度为45U/ml的酶,我想问,假设总体积为1ml,那么终浓度为45U/ml的酶岂不是每分钟能转化4.8μmol的底物?那么0.1μmol/mL的底物不就几秒钟就反应完了?那么怎么测定初速度?
2、可逆抑制剂:包括
a、竞争性抑制剂,抑制剂与底物竞争性结合酶反应中心,使Km增大,而Vmax不变 b、非竞争性抑制剂,酶与抑制剂结合后还能与底物结合,但活性降低,使Vmax减小,而Km不变
c、反竞争性抑制剂,酶只能与底物结合后才能与抑制剂结合,Vmax与Km都减小
可逆抑制剂可用透析等方法除去,使酶恢复作用
《血管紧张素转换酶抑制剂在心血管病中应用的中国专家共识》.PDF(242.69k)

