Literature DB >> 25368106

A rapid ceramide synthase activity using NBD-sphinganine and solid phase extraction.

Rotem Tidhar1, Kacee Sims2, Eden Rosenfeld-Gur1, Walter Shaw2, Anthony H Futerman1.   

Abstract

Ceramides are synthesized by six mammalian ceramide synthases (CerSs), each of which uses fatty acyl-CoAs of different chain lengths for N-acylation of the sphingoid long-chain base. We now describe a rapid and reliable CerS assay that uses a fluorescent N-[6-[(7-nitrobenzo-2-oxa-1,3-diazol-4-yl) (NBD) sphinganine substrate followed by separation of the NBD-lipid substrate and products using solid phase extraction (SPE) C18 chromatography. SPE chromatography is a quick and reliable alternative to TLC, and moreover, there is no degradation of either NBD-sphinganine or NBD-ceramide. We have optimized the assay for use with minimal amounts of protein in a minimal volume. This assay will prove useful for the analysis of CerS activity, which is of particular importance in light of the growing involvement of CerS in cell regulation and in the pathology of human diseases.
Copyright © 2015 by the American Society for Biochemistry and Molecular Biology, Inc.

Entities:  

Keywords:  N-acylation; ceramides; chromatography; enzymology; fatty acyl-CoA; in vitro assay; long-chain base; sphingolipids

Mesh：

Substances：

Year:  2014        PMID： 25368106      PMCID： PMC4274067          DOI： 10.1194/jlr.D052001

Source DB:  PubMed          Journal:  J Lipid Res        ISSN： 0022-2275            Impact factor:   5.922


  26 in total

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2.  Modulation of ceramide synthase activity via dimerization.

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Journal:  Adv Cancer Res       Date:  2013       Impact factor: 6.242

4.  Impaired epidermal ceramide synthesis causes autosomal recessive congenital ichthyosis and reveals the importance of ceramide acyl chain length.

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6.  Myristate-derived d16:0 sphingolipids constitute a cardiac sphingolipid pool with distinct synthetic routes and functional properties.

Authors:  Sarah Brice Russo; Rotem Tidhar; Anthony H Futerman; L Ashley Cowart
Journal:  J Biol Chem       Date:  2013-03-25       Impact factor: 5.157

7.  Two mammalian longevity assurance gene (LAG1) family members, trh1 and trh4, regulate dihydroceramide synthesis using different fatty acyl-CoA donors.

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Review 8.  Ceramide in cystic fibrosis.

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Journal:  Handb Exp Pharmacol       Date:  2013

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Authors:  Helena Van Overloop; Gerd Van der Hoeven; Paul P Van Veldhoven
Journal:  J Lipids       Date:  2012-07-26
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  13 in total

1.  Oxidative stress elicited by modifying the ceramide acyl chain length reduces the rate of clathrin-mediated endocytosis.

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Journal:  Anal Biochem       Date:  2019-03-24       Impact factor: 3.365

4.  Generation of a ceramide synthase 6 mouse lacking the DDRSDIE C-terminal motif.

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5.  Following the flux of long-chain bases through the sphingolipid pathway in vivo using mass spectrometry.

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Journal:  J Lipid Res       Date:  2016-03-14       Impact factor: 5.922

Review 6.  Sphingolipids as targets for treatment of fungal infections.

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Journal:  Future Med Chem       Date:  2016-08-09       Impact factor: 3.808

7.  Eleven residues determine the acyl chain specificity of ceramide synthases.

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8.  Characterizing a Common CERS2 Polymorphism in a Mouse Model of Metabolic Disease and in Subjects from the Utah CAD Study.

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Journal:  J Clin Endocrinol Metab       Date:  2021-07-13       Impact factor: 6.134

9.  The Role of Ceramide Synthases in the Pathogenicity of Cryptococcus neoformans.

Authors:  Mansa A Munshi; Justin M Gardin; Ashutosh Singh; Chiara Luberto; Robert Rieger; Tejas Bouklas; Bettina C Fries; Maurizio Del Poeta
Journal:  Cell Rep       Date:  2018-02-06       Impact factor: 9.423

10.  Effect of the sphingosine kinase 1 selective inhibitor, PF-543 on arterial and cardiac remodelling in a hypoxic model of pulmonary arterial hypertension.

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