Identication of Golgi-localized acyl transferases that palmitoylate and regulate endothelial nitric oxide synthase

被引:135
作者
Fernandez-Hernando, Carlos
Fukata, Masaki
Bernatchez, Pascal N.
Fukata, Yuko
Lin, Michelle I.
Bredt, David S.
Sessa, William C.
机构
[1] Yale Univ, Sch Med, Dept Pharmacol, New Haven, CT 06536 USA
[2] Yale Univ, Sch Med, Program Vascular Cell Signaling & Therapeut, Boyer Ctr Mol Med, New Haven, CT 06536 USA
[3] Natl Inst Longev Sci, Lab Genom & Proteom, Aichi 4748522, Japan
[4] Japan Sci & Technol Agcy, Precursory Res Embryon Sci & Technol, Kawaguchi, Saitama 3320012, Japan
[5] Eli Lilly & Co, Dept Integrat Biol, Indianapolis, IN 46285 USA
关键词
D O I
10.1083/jcb.200601051
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Lipid modi. cations mediate the subcellular localization and biological activity of many proteins, including endothelial nitric oxide synthase (eNOS). This enzyme resides on the cytoplasmic aspect of the Golgi apparatus and in caveolae and is dually acylated by both N-myristoylation and S-palmitoylation. Palmitoylation deficient mutants of eNOS release less nitric oxide ( NO). We identify enzymes that palmitoylate eNOS in vivo. Transfection of human embryonic kidney 293 cells with the complementary DNA (cDNA) for eNOS and 23 cDNA clones encoding the Asp-His-His-Cys motif (DHHC) palmitoyl transferase family members showed that five clones (2, 3, 7, 8, and 21) enhanced incorporation of [H-3]palmitate into eNOS. Human endothelial cells express all five of these enzymes, which colocalize with eNOS in the Golgi and plasma membrane and interact with eNOS. Importantly, inhibition of DHHC-21 palmitoyl transferase, but not DHHC-3, in human endothelial cells reduces eNOS palmitoylation, eNOS targeting, and stimulated NO production. Collectively, our data describe five new Golgi-targeted DHHC enzymes in human endothelial cells and suggest a regulatory role of DHHC-21 in governing eNOS localization and function.
引用
收藏
页码:369 / 377
页数:9
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