Identification and characterization of a novel human type II diacylglycerol kinase, DGK kappa.
Diacylglycerol kinase (DGK) plays an important role in signal transduction through modulating the balance between two signaling lipids, diacylglycerol and phosphatidic acid. Here we identified a tenth member of the DGK family designated DGK kappa. The kappa-isozyme (1271 amino acids, calculated molecular mass, 142 kDa) contains a pleckstrin homology domain, two cysteine-rich zinc finger-like structures, and a separated catalytic region as have been found commonly for the type II isozymes previously cloned (DGKdelta and DGKeta). The new DGK isozyme has additionally 33 tandem repeats of Glu-Pro-Ala-Pro at the N terminus. Reverse transcriptase-PCR showed that the DGK kappa mRNA is most abundant in the testis, and to a lesser extent in the placenta. DGK kappa, when expressed in HEK293 cells, was persistently localized at the plasma membrane even in the absence of cell stimuli. Deletion analysis revealed that the short C-terminal sequence (amino acid residues 1199-1268) is necessary and sufficient for the plasma membrane localization. Interestingly, DGK kappa, but not other type II DGKs, was specifically tyrosine-phosphorylated at Tyr78 through the Src family kinase pathway in H2O2-treated cells. Moreover, H2O2 selectively inhibited DGK kappa activity in a Src family kinase-independent manner, suggesting that the isozyme changes the balance of signaling lipids in the plasma membrane in response to oxidative stress. The expression patterns, subcellular distribution, and regulatory mechanisms of DGK kappa are distinct from those of DGKdelta and DGKeta despite high structural similarity, suggesting unique functions of the individual type II isozymes.
Imai
Shin-ichi
S
Department of Biochemistry, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo 060-8556, Japan.
Kai
Masahiro
M
Yasuda
Satoshi
S
Kanoh
Hideo
H
Sakane
Fumio
F
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000737:chemistry; D015153:Blotting, Western; D019556:COS Cells; D002384:Catalysis; D020134:Catalytic Domain; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003545:Cysteine / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D019852:Diacylglycerol Kinase / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D004075:Diglycerides / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D017353:Gene Deletion; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D015151:Immunoblotting; D047468:Immunoprecipitation; D008055:Lipids / Q000737:chemistry; D008297:Male; D008957:Models, Genetic; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D010712:Phosphatidic Acids / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D013552:Swine; D013737:Testis / Q000378:metabolism; D014018:Tissue Distribution; D014443:Tyrosine / Q000737:chemistry; D016335:Zinc Fingers
D001798:Blood Proteins; D018076:DNA, Complementary; D004075:Diglycerides; D008055:Lipids; D010712:Phosphatidic Acids; D010750:Phosphoproteins; D020033:Protein Isoforms; D012333:RNA, Messenger; C039077:platelet protein P47; D014443:Tyrosine; D006861:Hydrogen Peroxide; D019852:Diacylglycerol Kinase; D003545:Cysteine