Isolation and characterization of a cDNA encoding a Δ8 sphingolipid desaturase from Aquilegia vulgaris. (November 2002)

• Record Type: Journal Article
• Authors: Michaelson, L. V.; Longman, A. J.; Sayanova, O.; Stobart, A. K.; Napier, J. A.
• Is Part Of: Biochemical Society transactions. Volume 30:Number 6(2002)
• Page Start: 1073
• Page End: 1075
• Publication Date: 2002-11
• Subjects: cytochrome b5 fusion -- long-chain base -- sphingoid base; LCB, long-chain base; Biochemistry -- Congresses; 572
• DOI: 10.1042/bst0301073
• Languages: English
• ISSNs: 0300-5127

Abstract

Abstract : We have isolated a cDNA encoding the Δ 8 sphingolipid desaturase from the plant Aquilegia vulgaris L. via a PCR-based strategy using primers designed to target the conserved histidine box regions of microsomal desaturases. The function of the cDNA was confirmed by expression in the yeast, Saccharomyces cerevisiae. Analysis of the long-chain sphingoid bases as their dinitrophenyl derivatives by reverse-phase HPLC demonstrated the accumulation of cis - and trans -desaturated sphingoid bases which were not present in the wild-type yeast cells. The Δ 8 desaturated products co-eluted with known Δ 8 -desaturated phytosphingenine and the molecular mass of these products was confirmed by liquid chromatography-MS. The Δ 8 long-chain base desaturase was also able to desaturate dihydrosphingosine substrates. This is the first report of the functional characterization of an A. vulgaris gene product.