Abstract
Complementary collision-induced/electron capture dissociation Fourier-transform ion cyclotron resonance mass spectrometry was used to fully sequence the protein P2 myelin basic protein. It is an antigenic fatty-acid-binding protein that can induce experimental autoimmune neuritis: an animal model of Guillain–Barré syndrome, a disorder similar in etiology to multiple sclerosis. Neither the primary structure of the porcine variant, nor the fatty acids bound by the protein have been well established to date. A 1.8-Å crystal structure shows but a bound ligand could not be unequivocally identified. A protocol for ligand extraction from protein crystals has been developed with subsequent gas chromatography MS analysis allowing determination that oleic, stearic, and palmitic fatty acids are associated with the protein. The results provide unique and general evidence of the utility of mass spectrometry for characterizing proteins from natural sources and generating biochemical information that may facilitate attempts to elucidate the causes for disorders such as demyelination.
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Acknowledgments
We thank Stefan Svensson Gelius (Biovitrum, Sweden) for helpful discussion, Daniel Daley (Biochemistry Department, Stockholm University, Sweden) for technical and biochemical advice, and Maria Athanasiadou (Materials and Environmental Chemistry Department, Stockholm University, Sweden) for the chemical derivatization prior to GC/MS analysis. Research supported by grant 2007-6890 from the Swedish Research Council (JS).
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The last two addresses below are Dr. Jan Sedzik's present affiliations.
An erratum to this article can be found at http://dx.doi.org/10.1007/s00216-010-4204-8
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Maddalo, G., Shariatgorji, M., Adams, C.M. et al. Porcine P2 myelin protein primary structure and bound fatty acids determined by mass spectrometry. Anal Bioanal Chem 397, 1903–1910 (2010). https://doi.org/10.1007/s00216-010-3762-0
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DOI: https://doi.org/10.1007/s00216-010-3762-0