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. 2006 Dec 15;359(2):262-4.
doi: 10.1016/j.ab.2006.08.039. Epub 2006 Sep 22.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of cardiolipin extracted from detergent-solubilized mitochondrial electron transfer complexes

Tiffany McDonald-Marsh  1 , Christopher A Carroll, Neal C Robinson, Andrej Musatov
Affiliations

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of cardiolipin extracted from detergent-solubilized mitochondrial electron transfer complexes

Tiffany McDonald-Marsh et al. Anal Biochem. .
No abstract available

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Figures

Fig. 1
Fig. 1
Silicic acid HPLC purification of standard cardiolipin and cardiolipin extracted from dodecyl maltoside-solubilized cytochrome c oxidase or complex I. (A) Separation of mixed phospholipid standards (2 nmol of cardiolipin (CL), 4 nmol of phosphatidylethanolamine (PE), and 4 nmol of phosphatidylcholine (PC)). (B) HPLC analysis of phospholipids extracted from HiTrapQ-purified 1.0 nmol of cytochrome c oxidase. (C) HPLC analysis of phospholipids extracted from 1 nmol of isolated complex I. Detection of phospholipids elution was monitored at 203 nm with a Waters 2487 Dual λ Absorbance Detector.
Fig. 2
Fig. 2
MALDI-TOF/MS of standard cardiolipin and cardiolipin extracted from cytochrome c oxidase or complex I. (A) MALDI-TOF mass spectra of standard cardiolipin. Beef heart standard cardiolipin was obtained from Avanti Polar Lipids. (B) MALDI-TOF mass spectra of HPLC-isolated cardiolipin extracted from HiTrapQ-purified cytochrome c oxidase. (C) MALDI-TOF mass spectra of HPLC-isolated cardiolipin extracted from complex I. In each case cardiolipin was dissolved in chloroform and ∼15 pmol/μL spotted on the MALDI target using dihydroxybenzoic acid as the matrix. Each spectrum represents an average of 100 laser shots. Observed major peaks correspond to tetra-linoleoyl cardiolipin (1448 ± 1) modified by sodium ions.
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References

    1. Robinson NC. Functional binding of cardiolipin to cytochrome c oxidase. J. Bioenerg. Biomembr. 1993;25:153–163. - PubMed
    1. Iverson SL, Orrenius S. The cardiolipin-cytochrome c interaction and the mitochondrial regulation of apoptosis. Arch. Biochem. Biophys. 2004;423:37–46. - PubMed
    1. Musatov A. Contribution of peroxidized cardiolipin to inactivation of bovine heart cytochrome c oxidase. Free Radical Biol. Med. 2006;41:238–246. - PubMed
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    1. Weintraub ST, Musatov A, Carroll CA, Robinson NC. Analysis of mitochondrial electron transport complexes by MALDI-TOF mass spectrometry. A 50th American Society Mass Spectrometry Conference; Orlando, (USA). 2002.

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