
Overview:
RSK2 is a member of the RSK (ribosomal S6 kinase) family that consists of growth factor-regulated serine/threonine kinases. RSK2 has been shown to mediate growth factor signaling via RAS and MAPK leading to the induction of CREB serine-133 phosphorylation and activation of gene expression (1). Mutations in RSK2 have been shown to be responsible for Coffin-Lowry syndrome (CLS) which is a X-linked disorder characterized by severe psychomotor retardation, facial and digital dysmorphisms, and progressive skeletal deformations (2).
Gene Aliases:
RPS6KA3; HU-3; MAPKAPK1B; CLS; MRX19; ISPK-1; p90-RSK2; pp90RSK2; S6K-alpha3
Genbank Number:
NM_004586
References:
1. Xing, J. et al: Coupling of the RAS-MAPK pathway to gene activation by RSK2, a growth factor-regulated CREB kinase.Science. 1996 Aug 16;273(5277):959-63.2. Jacquot, S. et al: Mutation analysis of the RSK2 gene in Coffin-Lowry patients: extensive allelic heterogeneity and a high rate of de novo mutations. Am J Hum Genet. 1998 Dec;63(6):1631-40.
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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?
《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)
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

