Abstract
We report a multiple-stage ion-trap (IT) mass spectrometric approach with electrospray ionization (ESI) for structural characterization of the [M−2H+Na]− ion of cardiolipin (CL), a 1,3-bisphosphatidyl-sn-glycerol that consists of four fatty acyl chains and three glycerol backbones designated as A, B, and central glycerol, respectively (see Scheme 1). Following collisionally activated dissociation (CAD), the [M−2H+Na]− ions of CL yield two prominent fragment ions that arise from the differential losses of the diacylglycerol moieties containing A or B glycerol, respectively. The tentative assignment of the two phosphatidyl moieties attached to the 1′- or 3′-position of the central glycerol is based on the observation that the ions arising from loss of the diacylglycerol moiety containing glycerol B is more abundant than that containing glycerol A. The structures of the above two ions, including the identities of the fatty acyl substituents and the position of fatty acyl substituents on the glycerol backbones (glycerol A and B) are determined by MS3 experiments that give spectra comprising several sets of prominent ions informative for the structural assignment of the fatty acyl substituents on the glycerol A and glycerol B. This method permits the structures of CL in a mixture isolated from Escherichia coli, including species that consist of various isomers, to be unveiled in detail.
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Valianpour, F.; Wanders, R. J. A.; Barth, P. G.; Overmars, H.; van Gennip, A. H. Quantitative and Compositional Study of Cardiolipin in Platelets by Electrospray Ionization Mass Spectrometry: Application for the Identification of Barth Syndrome Patients. Clin. Chem. 2002, 48, 1390–1397.
Beckedorf, A. I.; Schaffer, C.; Messner, P.; Peter-Katalinic, J. Mapping and Sequencing of Cardiolipins from Geobacillus stearothermophilus NRS 2004/3a by Positive and Negative Ion Nano ESI-QTOF-MS and MS/MS. J. Mass Spectrom. 2002, 37, 1086–1094.
Lesnefsky, E. S.; Stoll, M. S. K.; Minkler, P. E.; Hoppel, C. L. Separation and Quantitation of Phospholipids and Lysophospholipids by High-Performance Liquid Chromatography. Anal. Biochem. 2000, 285, 246–254.
Peter-Katalinic, J.; Fischer, W. α-D-Glucopyranosyl-, D-alanyl-, and L-Lysylcardiolipin from Gram-Positive Bacteria: Analysis by Fast Bombardment Mass Spectrometry. J. Lipid Res. 1998, 39, 2286–2292.
Hsu, F. F.; Turk, J.; Rhoades, E. R.; Russell, D. G.; Shi, Y.; Groisman, E. A. Structural Characterization of Cardiolipin by Tandem Quadrupole and Multiple-Stage Quadrupole Ion-Trap Mass Spectrometry with Electrospray Ionization. J. Am. Soc. Mass Spectrom. 2005, 16, 491–504.
LeCocq, J.; Ballou, C. E. On the Structure of Cardiolipin. Biochemistry 1964, 155, 976–980.
Powell, G. L.; Jacobus, J. The Nonequivalence of the Phosphorus Atoms in Cardiolipin. Biochemistry 1974, 13, 4024–4026.
Hsu, F. F.; Turk, J. Characterization of Phosphatidylinositol, Phosphatidylinositol-4-Phosphate, and Phosphatidylinositol-4,5-Bisphosphate by Electrospray Tandem Mass Spectrometry: A Mechanistic Study. J. Am. Soc. Mass Spectrom. 2000, 11, 986–999.
Hsu, F. F.; Turk, J. Charge-Driven Fragmentation Processes in Diacyl Glycerophosphatidic Acids Upon Low-Energy Collisional Activation. A Mechanistic Proposal. J. Am. Soc. Mass Spectrom. 2000, 11, 797–803.
Hsu, F. F.; Turk, J. Charge-Driven and Charge-Remote Fragmentation Processes in Diacyl Glycerophosphoethanolamine Upon Low-Energy Collisional Activation. A Mechanistic Proposal. J. Am. Soc. Mass Spectrom. 2000, 11, 892–899.
Henderson, T. O.; Glonek, T.; Myers, T. C. Phosphorus-31 Nuclear Magnetic Resonance Spectroscopy of Phospholipids. Biochemistry 1974, 13, 623–628.
Kates, M.; Syz, J. Y.; Gosser, D.; Haines, T. H. pH-Dissociation Characteristics of Cardiolipin and its 2′-deoxy Analog. Lipids. 1993, 28, 877–882.
Schlame, M.; Brody, S.; Hostetler, K. Y. Mitochondrial Cardiolipin in Diverse Eukaryotes. Comparison of Biosynthetic Reactions and Molecular Acyl Species. Eur. J. Biochem.2127271993735
Schlame, M.; Otten, D. Analysis of Cardiolipin Molecular Species by High-Performance Liquid Chromatography of Its Derivative 1,3-Bisphosphatidyl-2-Benzoyl-sn-Glycerol Dimethyl Ester. Anal. Biochem. 1991, 195, 290–295.
Hirschberg, C. B.; Kennedy, E. P. Mechanism of the Enzymatic Synthesis of Cardiolipin in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 1972, 69, 648–651.
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Published oline January 25, 2006
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Hsu, F.F., Turk, J. Characterization of cardiolipin from Escherichia coli by electrospray ionization with multiple stage quadrupole ion-trap mass spectrometric analysis of [M−2H+Na]− ions. The official journal of The American Society for Mass Spectrometry 17, 420–429 (2006). https://doi.org/10.1016/j.jasms.2005.11.019
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DOI: https://doi.org/10.1016/j.jasms.2005.11.019