IDENTIFICATION OF PHOSPHORYLATION SITES ON MURINE NUCLEAR LAMIN-C BY RP-HPLC AND MICROSEQUENCING
IDENTIFICATION OF PHOSPHORYLATION SITES ON MURINE NUCLEAR LAMIN-C BY RP-HPLC AND MICROSEQUENCING
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DOI：
10.1016/0014-5793(91)80868-4
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发表时间：
1991-11-04
期刊：
影响因子：
3.5
通讯作者：
JOST, E
中科院分区：
文献类型：
--
作者：
EGGERT, M;RADOMSKI, N;JOST, E
从艾氏腹水肿瘤细胞中获得分离的间期核纤层蛋白 C，用 Lys-C 内切蛋白酶消化，所得肽通过反相 HPLC 分离，并进行微测序，以鉴定间期磷酸化位点以及体外磷酸化位点，分别由 cdc2 激酶、蛋白激酶 C (PKC) 和蛋白激酶 A (PKA) 进行。 核纤层蛋白 C 显示 Ser392 和 Ser409 部分磷酸化，间期可能还有 Ser407。 Ser390 和 Ser392 处的 cdc2 激酶以及 Ser572 处的 PKC 导致磷酸化增加。 含有 3 种激酶共有序列的 N 端肽 (aa 1-32) 被 cdc2 激酶、PKC 和 PKA 磷酸化。 序列数据表明，通过核纤层修饰的多个分子开关控制着细胞周期中核骨架的动态行为。
Isolated interphase lamin C, obtained from Ehrlich ascites tumor cells, was digested by Lys-C endoproteinase, the resulting peptides separated by reversed-phase HPLC and subjected to microsequencing in order to identify phosphorylation sites in interphase and following phosphorylation in vitro by cdc2-kinase, protein kinase C (PKC) and protein kinase A (PKA), respectively. Nuclear lamin C showed partial phosphorylation of Ser392 and Ser409, and possibly Ser407 in interphase. Phosphorylation was increased in response to cdc2-kinase at Ser390 and Ser392 and to PKC at Ser572. The N-terminal peptide (aa 1-32) containing consensus sequences for the 3 kinases was phosphorylated by cdc2-kinase, PKC and PKA. The sequence data suggests that multiple molecular switches via lamina modification control the dynamic behaviour of the nucleoskeleton during the cell cycle.