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Characterization of PSKH1, a novel human protein serine kinase with centrosomal, golgi, and nuclear localization.

机译:PSKH1的特征,一种具有中心体,高尔基体和核定位的新型人类蛋白丝氨酸激酶。

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We report here the characterization of PSKH1, a novel human protein serine kinase with multiple intracellular localizations. The gene consists of three exons distributed over 35 kb of genomic DNA in region 16q22.1. The 3.4-kb cDNA predicts a protein of 424 amino acids with a calculated molecular mass of 48.1 kDa and pI of 9.6. PSKH1 is expressed in all tissues and cell lines tested as shown by Northern blots, with the highest level of abundance in testis. PSKH1 displays the highest level of similarity with rat CaM kinase I (50. 2%) over 259 amino acids in the conserved catalytic region, but lacks significant homology with proteins in the database outside the catalytic core. Polyclonal antibodies have been raised, and indirect immunofluorescence microscopy of untransfected COS-1 cells suggests that PSKH1 is localized in the Brefeldin A-sensitive Golgi compartment, at centrosomes, in the nucleus with a somewhat speckle-like presence, and more diffusely in the cytoplasm. The presence in the centrosome appears to be enhanced during osmotic stress. Immunoisolated PSKH1 does not phosphorylate any of the common kinase substrates in vitro, but autophosphorylates exclusively serines within its COOH-terminal region in an intermolecular fashion. Furthermore, autophosphorylation activity is repressed upon addition of Ca(2+)/CaM, suggesting that PSKH1 activity depends on Ca(2+) concentration in vivo. Copyright 2000 Academic Press.
机译:我们在这里报告PSKH1的表征,一种具有多个细胞内定位的新型人类蛋白丝氨酸激酶。该基因由分布在16q22.1区35 kb基因组DNA上的三个外显子组成。 3.4 kb cDNA可以预测424个氨基酸的蛋白质,计算分子量为48.1 kDa,pI为9.6。 PSKH1在所有组织和细胞系中均表达,如Northern印迹法所示,睾丸中的丰度最高。 PSKH1在保守的催化区域中与259个氨基酸处的大鼠CaM激酶I(50. 2%)具有最高的相似性,但与催化核心外部的数据库中的蛋白质缺乏显着的同源性。已经产生了多克隆抗体,未转染的COS-1细胞的间接免疫荧光显微镜检查表明,PSKH1位于布雷菲德菌素A敏感的高尔基体中,在中心体中,在核中有一些斑点样的存在,并且在细胞质中更弥散。在渗透胁迫期间似乎增加了中心体的存在。免疫分离的PSKH1不会在体外磷酸化任何常见的激酶底物,但会以分子间方式仅在其COOH末端区域内自磷酸化丝氨酸。此外,添加Ca(2 +)/ CaM后,自身磷酸化活性受到抑制,这表明PSKH1活性取决于体内Ca(2+)的浓度。版权所有2000学术出版社。

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