Kinetic characterization of mammalian ceramide synthases: Determination of Km values towards sphinganine Academic Article uri icon

Overview

MeSH Major

  • Oxidoreductases
  • Sphingosine

abstract

  • 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 K(m) value of each CerS towards sphinganine. Remarkably, the K(m) values towards sphinganine are all similar, ranging from 2 to 5microM, 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.

publication date

  • November 13, 2007

Research

keywords

  • Academic Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1016/j.febslet.2007.10.018

PubMed ID

  • 17977534

Additional Document Info

start page

  • 5289

end page

  • 94

volume

  • 581

number

  • 27