Kinetic characterization of mammalian ceramide synthases:: Determination of Km values towards sphinganine

被引：70

作者：

Lahiri, Sujoy ^{[1
]}

Lee, Hyunmi ^{[2
]}

Mesicek, Judith ^{[2
]}

Fuks, Zvi ^{[3
]}

Haimovitz-Friedman, Adriana ^{[3
]}

Kolesnick, Richard N. ^{[2
]}

Futerman, Anthony H. ^{[1
]}

机构：

[1] Weizmann Inst Sci, Dept Biol Chem, IL-76100 Rehovot, Israel

[2] Mem Sloan Kettering Canc Ctr, Lab Signal Transduct, New York, NY 10021 USA

[3] Mem Sloan Kettering Canc Ctr, Dept Radiat Oncol, New York, NY 10021 USA

来源：

FEBS LETTERS | 2007年 / 581卷 / 27期

关键词：

lipid; sphingolipid; ceramide; acyl CoA; sphinganine; assay;

D O I：

10.1016/j.febslet.2007.10.018

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Ceramide is a key metabolite in the pathway of sphingolipid biosynthesis. In mammals, ceramide is synthesized by N-acylation of a sphingoid long-chain base by a family of ceramide synthases (CerS), each of which displays a high specificity towards acyl CoAs of different chain lengths. We now optimize a previously-described assay for measuring CerS activity for use upon over-expression of mammalian CerS, and using these conditions, establish the Km value of each CerS towards sphinganine. Remarkably, the Km values towards sphinganine are all similar, ranging from 2 to 5 mu M, even for CerS proteins that are able to use more than one acyl CoA for ceramide synthesis (i.e. CerS4). The availability of this assay will permit further accurate characterization of the kinetic parameters of mammalian CerS proteins. (C) 2007 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.

引用

页码：5289 / 5294

页数：6

共 28 条

[1]

BLIGH EG, 1959, CAN J BIOCHEM PHYS, V37, P911

[2] CERAMIDE SYNTHASE MEDIATES DAUNORUBICIN-INDUCED APOPTOSIS - AN ALTERNATIVE MECHANISM FOR GENERATING DEATH SIGNALS [J].