CDC2-LIKE KINASES IN NORMAL AND MALIGNANT CELLS

最近的研究表明，存在一个哺乳动物蛋白质家族， 结构和功能上与酵母细胞周期相关的激酶 调节激酶cdc-2。至少有5个cdc 2家族成员，cdk 1 通过CDK 5，已经在哺乳动物细胞中被鉴定。本实验室 专注于研究cdk 5的特性，与 cdk 1或cdk 2，cdk 5在细胞终末和终末都有高水平表达， 分化的神经元和许多培养的细胞系。我们已经生成了一个 特异性针对CDK 5的抗肽抗血清， 免疫沉淀来自哺乳动物细胞提取物CDK 5激酶，并结合 在Western印迹研究中特异性结合CDK 5。使用这种抗血清， 我们已经确定，培养的人类肿瘤细胞表达酶促 活性CDK 5激酶我们目前正试图重建活跃的 CDK 5酶在杆状病毒中表达后的体外试验和随后的 纯化，这将允许识别和表达关键的 调节活性的调节分子。此外，我们还克隆了 并鉴定了编码果蝇同源物的cDNA和基因组克隆， 并将该基因定位于2号染色体上的一条带， 提供所需的信息来产生遗传缺陷的苍蝇， 在cdk 5基因中。

Recent studies have shown that there exists a family of mammalian protein kinases structurally and functionally related to the yeast cell cycle regulatory kinase cdc-2. At least five members of the cdc2 family, cdk1 through cdk5, have been identified in mammalian cells. Our laboratory is focused on studying the properties of cdk5 which, in contrast to either cdk 1 or cdk2, cdk 5 is expressed at high levels both in terminally differentiated neurons and many cultured cell lines. We have generated an antipeptide antiserum specific for cdk 5, which can specifically immunoprecipitate cdk 5 kinase from mammalian cell extracts, and binds specifically to cdk 5 in Western blotting studies. Using this antiserum, we have established that cultured human tumor cells express enzymatically active cdk 5 kinase. We are currently attempting to reconstitute active cdk5 enzyme in vitro after expression in baculovirus and subsequent purification, which will allow identification and expression of key regulatory molecules that modulate activity. In addition, we have cloned and characterized cDNA and genomic clones encoding the Drosophila homolog for mammalian cdk5 and mapped the gene to a band on chromosome 2, thereby providing the information needed to generate flies genetically deficient in the cdk5 gene.