Functional characterization of human sphingosine kinase‐1

FEBS Letters

Kohama, Takafumi¹  Sugiura, Masako¹  Kono, Keita¹  Lacana’, Emanuela³  Milstien, Sheldon²  Spiegel, Sarah³  Nava, Victor E.³  Liu, Hong³  Poulton, Samantha³ 

[1] Pharmacology and Molecular Biology Research Laboratories, Sankyo Co., Ltd., Tokyo 140-8710, Japan;LCMR, NIMH, Bethesda, MD 20892, USA;Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, 353 Basic Science Building, 3900 Reservoir Road NW, Washington, DC 20007, USA

关键词: Human sphingosine kinase;    Sphingosine 1-phosphate;    BSA;    bovine serum albumin;    DMS;    N;    N-dimethylsphingosine;    DHS;    D;    L-threo-dihydrosphingosine;    SPHK;    sphingosine kinase;    SPP;    sphingosine 1-phosphate;    PCR;    polymerase chain reaction;    RACE;    rapid amplification of cDNA ends;   

DOI  :  10.1016/S0014-5793(00)01510-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Sphingosine kinase catalyzes the phosphorylation of sphingosine to form sphingosine 1-phosphate (SPP), a novel lipid mediator with both intra- and extracellular functions. Based on sequence identity to murine sphingosine kinase (mSPHK1a), we cloned and characterized the first human sphingosine kinase (hSPHK1). The open reading frame of hSPHK1 encodes a 384 amino acid protein with 85% identity and 92% similarity to mSPHK1a at the amino acid level. Similar to mSPHK1a, when HEK293 cells were transfected with hSPHK1, there were marked increases in sphingosine kinase activity resulting in elevated SPP levels. hSPHK1 also specifically phosphorylated D-erythro-sphingosine and to a lesser extent sphinganine, but not other lipids, such as D,L-threo-dihydrosphingosine, N,N-dimethylsphingosine, diacylglycerol, ceramide, or phosphatidylinositol. Northern analysis revealed that hSPHK1 was widely expressed with highest levels in adult liver, kidney, heart and skeletal muscle. Thus, hSPHK1 belongs to a highly conserved unique lipid kinase family that regulates diverse biological functions.

【 授权许可】

Unknown