A novel high-molecular mass mammalian ethanolamine kinase.
The present report describes, for the first time, a mammalian kinase highly specific for ethanolamine (Km = 41 microM). Ethanolamine kinase catalyzes the initial step in the CDP-ethanolamine pathway for phosphatidylethanolamine synthesis. Although plant and protozoan kinases are known to exhibit remarkable specificity for ethanolamine, no equivalent mammalian kinase has been characterized previously. The easily shocked (eas) mutant of Drosophila has been characterized and found to lack ethanolamine kinase activity while still possessing normal choline kinase activity (Pavlidis, P., Ramaswami, M., and Tanouye, M.A. (1994) Cell 79, 23-33). The gene compensating this mutation encodes a sequence resembling that of choline kinase. In the present report, the eas gene product, expressed in Escherichia coli as fusion protein, is found to have highly specific ethanolamine kinase activity. Anti-eas antibody revealed a protein with a molecular mass of approximately 86 kDa in rat tissues and human HeLa cells by Western blotting, but did not bind rat choline kinase isozymes. Rat liver kinase activity specific for ethanolamine was separated chromatographically and by an immunological procedure using anti-choline kinase antibody, associated with the 86 kDa protein, and immunoprecipitated by anti-eas antibody. This 86 kDa protein is characterized as ethanolamine kinase. Relations with previously reported kinases are discussed.
Uchida
T
T
Department of Biochemistry, Gunma University School of Medicine, Maebashi, Japan. tuchida@sb.gunma-u.ac.jp
D000818:Animals; D019856:Ethanolamine / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008297:Male; D008970:Molecular Weight; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
D011993:Recombinant Fusion Proteins; D019856:Ethanolamine; D017853:Phosphotransferases (Alcohol Group Acceptor); C020125:ethanolamine kinase