
Overview:
ERN1 or endoplasmic reticulum to nucleus signaling 1 is the ER to nucleus signalling 1 protein, a human homologue of the yeast Ire1 gene product which possesses intrinsic kinase activity and an endoribonuclease activity and it is important in endoplasmic reticulum-based stress signals. ERN1 controls IRE1 proteolysis in mammalian cells (1). The activation of ERN1 through oligomerization expands the mechanistic repertoire of kinase-based signaling receptors. ERNI has a critical function in extraembryonic cells that is essential for fetal viability (2).
Gene Aliases:
hIRE1p; IRE1; IRE1A; IRE1P
Genbank Number:
NM_001433
References:
1. Niwa, M.et.al: A role for presenilin-1 in nuclear accumulation of Ire1 fragments and induction of the mammalian unfolded protein response. Cell 99: 691-702, 1999.2. Iwawaki, T.et.al: Function of IRE1 alpha in the placenta is essential for placental development and embryonic viability. Proc. Nat. Acad. Sci. 106: 16657-16662, 2009.
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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?

