Improved specificity of cardiolipin peroxidation by soybean lipoxygenase: a liquid chromatography - electrospray ionization mass spectrometry investigation.
Peroxidation catalysed by Soybean Lypoxigenase was performed on tetralinoleyl-cardiolipin with the aim of generating selectively oxidized products, to be used subsequently as standards for studies on cardiolipin oxidation. The reaction products were characterized by LC-ESI-MS and MS/MS, and the process was found to link a hydroperoxylic group on one or more linoleic chains of cardiolipin, up to a total of four groups per molecule. Interestingly, the incidence of other oxidized products, like those arising from multiple hydroxylation or mixed hydroxylation-hydroperoxydation, previously observed after the chemical oxidation of the same cardiolipin, was found to be negligible. Moreover, evidences for the presence of the hydroperoxylic group(s) almost exclusively on carbon 13 of the linoleic chain(s) were obtained by MS/MS measurements. The enzymatic approach, integrated with a preparative separation step, which could be developed by adapting the chromatographic conditions adopted in the present work for analytical purposes, represents a promising strategy for the synthesis of highly specific mono- or multi-peroxidated derivatives of cardiolipins.
Dipartimento di Chimica, Università degli Studi di Bari, Via E. Orabona 4, 70126, Bari, Italy. losito@chimica.uniba.it
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D000818:Animals; D002308:Cardiolipins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015227:Lipid Peroxidation; D008084:Lipoxygenase / Q000737:chemistry / Q000378:metabolism*; D013025:Glycine max / Q000201:enzymology*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods
D002308:Cardiolipins; D008084:Lipoxygenase