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Journal of Lipid Research, Vol. 50, 1120-1132, June 2009
Copyright © 2009 by American Society for Biochemistry and Molecular Biology

A shotgun lipidomics approach in Sinorhizobium meliloti as a tool in functional genomics

Libia Saborido Basconcillo^*, Rahat Zaheer, Turlough M. Finan and Brian E. McCarry^1,*

^* Department of Chemistry, McMaster University, Hamilton, Ontario, Canada
Department of Biology, McMaster University, Hamilton, Ontario, Canada

Published, JLR Papers in Press, December 18, 2008.

¹ To whom correspondence should be addressed. e-mail: mccarry{at}mcmaster.ca

A shotgun lipidomics approach that allowed the analysis of eight lipid classes directly from crude extracts of the soil bacterium Sinorhizobium meliloti is presented. New MS-MS transitions are reported for the analysis of monomethylphosphatidylethanolamines, dimethylphosphatidylethanolamines, and three bacterial non-phosphorus-containing lipid classes [sulfoquinovosyldiacylglycerols, ornithines, and diacylglyceryl-(N,N,N-trimethyl)-homoserines]. Unique MS-MS transitions allowed the analysis of isomeric species from various lipid classes without chromatography. Analyses required small sample amounts and minimal preparation; thus, this methodology has excellent potential to be used as a screening tool for the analysis of large numbers of samples in functional genomics studies. FA distributions within lipid classes of S. meliloti are described for the first time, and the relative positions of fatty acyl substituents (sn-1, sn-2) in phospholipids are presented. FA distributions in diacylglyceryl-(N,N,N-trimethyl)-homoserines were identical to those of phospholipids, indicating a common biosynthetic origin for these lipids. The method was applied to the analysis of mutants deficient in the PhoB regulator protein. Increased lipid cyclopropanation was observed in PhoB-deficient mutants under P_i starvation.

Supplementary key words electrospray ionization • tandem mass spectrometry • sulfoquinovosyldiacylglycerol • diacylglyceryl-(N,N,N-trimethyl)-homoserine • ornithine • P_i limitation • PhoB regulator

Abbreviations: CE, collision energy; CID, collision-induced dissociation; CL, cardiolipin; DMPE, dimethylphosphatidylethanolamine; ESI-MS, electrospray mass spectrometry; FAME, fatty acid methyl ester; HPTLC, high-performance TLC; MMPE, monomethylphosphatidylethanolamine; NP, normal phase; OL, ornithine lipid; PC, phosphatidylcholine; PCA, principal components analysis; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PHB, poly(3-hydroxybutyrate); P_i, inorganic phosphate; RP, reverse phase; SL, sulfoquinovosyldiacylglycerol; TMHS, 1,2-diacylglyceryl-3-O-4'-(N,N,N-trimethyl)-homoserine lipid

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