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Cloning and preliminary characterization of a 105 kDa protein with an N-terminal kinase-like domain

机译:具有N端激酶样结构域的105 kDa蛋白的克隆和初步表征

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摘要

In the course of searching for proteins that interact with protein kinase B in 3T3-L1 adipocytes, we isolated a 105 kDa protein from 3T3-L1 adipocytes. Peptides sequenced from the protein were found to be present in several expressed sequence tags. A cDNA containing one of these expressed sequence tags was sequenced and appears to contain the entire coding region. Computer analysis revealed a potential protein kinase domain at the N-terminus; however, the first subdomain and several invariant residues characteristic of protein kinases are absent. An antibody was raised against a peptide from the 105 kDa protein. By immunoblotting, it was found that the protein was widely expressed in mouse tissues, and concentrated in the cytosol and low density microsome fractions of 3T3-L1 adipocytes.
机译:在寻找与3T3-L1脂肪细胞中的蛋白激酶B相互作用的蛋白质的过程中,我们从3T3-L1脂肪细胞中分离了105 kDa的蛋白质。发现从蛋白质测序的肽存在于几个表达的序列标签中。对包含这些表达的序列标签之一的cDNA进行测序,似乎包含整个编码区。计算机分析表明在N端有潜在的蛋白激酶结构域。但是,缺少蛋白激酶的第一个亚结构域和几个不变的残基。产生了针对来自105 kDa蛋白的肽的抗体。通过免疫印迹,发现该蛋白在小鼠组织中广泛表达,并集中在3T3-L1脂肪细胞的细胞溶质和低密度微粒体级分中。

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