Porcine P2 myelin protein primary structure and bound fatty acids determined by mass spectrometry.
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.
Department of Analytical Chemistry, Stockholm University, Svante Arrhenius väg 16, 10691, Stockholm, Sweden.
Gianluca
G
Maddalo
Mohammadreza
M
Shariatgorji
Christopher M
CM
Adams
Eva
E
Fung
Ulrika
U
Nilsson
Roman A
RA
Zubarev
Jan
J
Sedzik
Leopold L
LL
Ilag
D000595:Amino Acid Sequence; D000818:Animals; D003460:Crystallization; D050556:Fatty Acid-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005227:Fatty Acids / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D018996:Myelin P2 Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D016415:Sequence Alignment; D013552:Swine
D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D018996:Myelin P2 Protein