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Porcine P2 myelin protein primary structure and bound fatty acids determined by mass spectrometry

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An Erratum to this article was published on 05 October 2010

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.

FT-ICR MS/MS spectrum (left) of porcine myelin P2 protein (green) and GC profile (right) of associated lipids extracted/identified from protein crystals by GC-MS. (Note: Ribbon diagram was generated by Rasmol based on PDB file 1YIV. Crystals depicted are not of the sample used.)

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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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Authors and Affiliations

  1. Department of Analytical Chemistry, Stockholm University, Svante Arrhenius väg 16, 10691, Stockholm, Sweden

    Gianluca Maddalo, Mohammadreza Shariatgorji, Ulrika Nilsson & Leopold L. Ilag

  2. Institute for Cell and Molecular Biology, Uppsala University, 75124, Uppsala, Sweden

    Christopher M. Adams, Eva Fung & Roman A. Zubarev

  3. Division of Molecular Biometry, Institute for Medicinal Biochemistry and Biophysics, Karolinska Institutet, Scheeles väg 2, 17177, Stockholm, Sweden

    Christopher M. Adams, Eva Fung & Roman A. Zubarev

  4. Department of Neurobiology, Health Care and Society, Protein Crystallization Facility, Karolinska Institute, NOVUM, 14186, Stockholm, Sweden

    Jan Sedzik

  5. Department of Chemical Engineering and Technology, Protein Crystallization Facility, Royal Institute of Technology, KTH, Teknikringen 28, 10044, Stockholm, Sweden

    Jan Sedzik

  6. Division of Neurobiology and Bioinformatic, National Institute of Physiological Science, Nishigonaka Myadaiji, Okazaki, 444-8585, Aichi, Japan

    Jan Sedzik

  7. Department of Applied Chemistry, School of Engineering 7-3-1, The University of Tokyo, Hongo, Bunkyo, 113-8656, Tokyo, Japan

    Jan Sedzik

Authors
  1. Gianluca Maddalo
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  2. Mohammadreza Shariatgorji
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  3. Christopher M. Adams
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  4. Eva Fung
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  7. Jan Sedzik
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  8. Leopold L. Ilag
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Corresponding author

Correspondence to Leopold L. Ilag.

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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

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