Overview:
TLK2 or Tousled-like kinase 2, is a nuclear serine/threonine kinases that is thought to be involved in the regulation of chromatin assembly (1). In addition, TLK2 is also involved in other cellular processes such as intracellular signaling cascade, protein amino acid phosphorylation, cell cycle, chromatin modification and in response to DNA damage stimulus. TLK2 displays maximal activity during S phase of the cell cycle where it appears to be regulated by cell-cycle-dependent phosphorylation. Inhibition of DNA replication causes a rapid loss of TLK2 activity, indicating that TLK2 function is tightly linked to ongoing DNA replication.
Gene Aliases:
MGC44450; PKU-ALPHA
Genbank Number:
NM_006852
References:
1.Yamakawa, A. et al: cDNA cloning and chromosomal mapping of genes encoding novel protein kinases termed PKU-alpha and PKU-beta, which have nuclear localization signal. Gene,1998; 202 (1-2): 193–201.2.Sillje, H. H. W. et al: Mammalian homologues of the plant Tousled gene code for cell-cycle-regulated kinases with maximal activities linked to ongoing DNA replication. EMBO J. 18: 5691-5702, 1999.
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《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)
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
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
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？
