Nishiura H, Tamura K, Morimoto Y, Imai H
Department of Biology, Faculty of Science, Konan University, Kobe 658-8501, Japan.
Biochem Soc Trans. 2000 Dec;28(6):747-8.
Sphingolipid long-chain base (LCB) kinase catalyses the phosphorylation of sphingolipid LCB to form LCB 1-phosphate. Based on sequence identity to a murine sphingosine kinase (murine SPHK1a), we isolated and characterized a LCB kinase-like cDNA in Arabidopsis thaliana. The deduced amino acid sequence of the homologous cDNA shows several regions that are highly conserved in LCB kinases from mouse, yeast, human and Caenorhabditis elegans. These regions are not similar to those of other known kinase families. For a functional identification, the homologous cDNA from A. thaliana was expressed in Escherichia coli, and LCB kinase activity was measured. The recombinant AtLcbk1 protein was found to utilize ATP and sphinganine. These results indicate the first identification of a gene coding for a LCB kinase in plants.
鞘脂长链碱基(LCB)激酶催化鞘脂长链碱基磷酸化形成1-磷酸长链碱基。基于与小鼠鞘氨醇激酶(小鼠SPHK1a)的序列同一性,我们在拟南芥中分离并鉴定了一个类似LCB激酶的cDNA。同源cDNA推导的氨基酸序列显示出几个在小鼠、酵母、人类和秀丽隐杆线虫的LCB激酶中高度保守的区域。这些区域与其他已知激酶家族的区域不相似。为了进行功能鉴定,将来自拟南芥的同源cDNA在大肠杆菌中表达,并测量LCB激酶活性。发现重组AtLcbk1蛋白利用ATP和鞘氨醇。这些结果表明首次在植物中鉴定出编码LCB激酶的基因。