Activation of PKN, a novel 120-kDa protein kinase with leucine zipper-like sequences, by unsaturated fatty acids and by limited proteolysis.
Activation of PKN, a novel 120-kDa protein kinase with leucine zipper-like sequences, by unsaturated fatty acids and by limited proteolysis.
复制标题
PKN 是一种新型 120 kDa 蛋白激酶，具有亮氨酸拉链样序列，通过不饱和脂肪酸和有限的蛋白水解作用进行激活。
DOI：
10.1006/bbrc.1994.2466
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发表时间：
1994
影响因子：
3.1
通讯作者：
Y. Ono
中科院分区：
文献类型：
--
作者：
H. Mukai;M. Kitagawa;H. Shibata;H. Takanaga;K. Mori;M. Shimakawa;M. Miyahara;K. Hirao;Y. Ono
PKN是一种新型蛋白激酶，具有与PKC家族同源的催化结构域和独特的氨基末端亮氨酸拉链状序列，从COS7细胞中纯化了部分编码人类PKN的cDNA构建体，用于酶学表征。使用含有基于PKC假底物位点的合成肽的丝氨酸作为磷酸盐受体，从部分纯化的PKN中估计的激酶活性不受Ca2+/磷脂酰丝氨酸/二油酸的刺激，但被40微克的不饱和脂肪酸（如花生四烯酸、亚油酸和油酸）激活数倍至数十倍。使用抗pkn抗血清的免疫沉淀的自磷酸化也被各种不饱和脂肪酸刺激。胰蛋白酶对PKN的有限蛋白水解诱导了几乎独立于花生四烯酸的肽激酶活性的增强。
PKN, a novel protein kinase with catalytic domain homologous to PKC family and unique amino terminal leucine zipper-like sequences, was purified partially from COS7 cells transfected with the cDNA construct encoding human PKN for enzymatic characterization of the enzyme. Using serine containing synthetic peptides based on PKC pseudosubstrate sites as the phosphate acceptors, kinase activities estimated from partially purified PKN were not stimulated by Ca2+/phosphatidylserine/diolein but were activated several-fold to several tens-fold by 40 microM unsaturated fatty acids, such as arachidonic acid, linoleic acid, and oleic acid. Autophosphorylation of the immunoprecipitates using anti-PKN antiserum was also stimulated by various unsaturated fatty acids. Limited proteolysis of PKN with trypsin induced an enhancement of the peptide kinase activity that was almost independent of arachidonic acid.