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

Anal Bioanal Chem. 2010 Jul;397(5):1903-10. doi: 10.1007/s00216-010-3762-0. Epub 2010 May 30.

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-A 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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Crystallization
  • Fatty Acid-Binding Proteins / chemistry*
  • Fatty Acid-Binding Proteins / genetics
  • Fatty Acid-Binding Proteins / metabolism
  • Fatty Acids / chemistry*
  • Fatty Acids / metabolism
  • Humans
  • Mass Spectrometry / methods*
  • Molecular Conformation
  • Molecular Sequence Data
  • Myelin P2 Protein / chemistry*
  • Myelin P2 Protein / genetics
  • Myelin P2 Protein / metabolism
  • Protein Binding
  • Sequence Alignment
  • Swine

Substances

  • Fatty Acid-Binding Proteins
  • Fatty Acids
  • Myelin P2 Protein