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

• Published in: Analytical and bioanalytical chemistry Vol. 397; no. 5; pp. 1903 - 1910
• Main Authors: Maddalo, Gianluca, Shariatgorji, Mohammadreza, Adams, Christopher M, Fung, Eva, Nilsson, Ulrika, Zubarev, Roman A, Sedzik, Jan, Ilag, Leopold L
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01-07-2010; Springer-Verlag; Springer
• Subjects: Amino Acid Sequence; Amino acids; Analytical Chemistry; Analytisk kemi; Animals; Biochemistry; Biokemi; Biologi; Biological and medical sciences; Biology; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatographic methods and physical methods associated with chromatography; CID; Crystalline structure; Crystallization; Crystals; De novo sequencing; ECD; Exact sciences and technology; Fatty Acid-Binding Proteins - chemistry; Fatty Acid-Binding Proteins - genetics; Fatty Acid-Binding Proteins - metabolism; Fatty acids; Fatty Acids - chemistry; Fatty Acids - metabolism; Food Science; Fundamental and applied biological sciences. Psychology; Gas chromatographic methods; GC/MS; Guillain-Barre syndrome; Humans; Kemi; Laboratory Medicine; Lipid identification; Mass spectrometry; Mass Spectrometry - methods; Mathematical analysis; MEDICIN; Medicin och hälsovetenskap; MEDICINE; Molecular biophysics; Molecular Conformation; Molecular Sequence Data; Monitoring/Environmental Analysis; Myelin; Myelin P2 Protein - chemistry; Myelin P2 Protein - genetics; Myelin P2 Protein - metabolism; Myelin proteins; NATURAL SCIENCES; NATURVETENSKAP; Original Paper; Protein Binding; Proteins; Sequence Alignment; Spectrometric and optical methods; Spectrum analysis; Structure; Structure in molecular biology; Swine; Lipid identification; De novo sequencing; GC/MS; Myelin proteins; ECD; CID; Animal model; Fourier transform spectroscopy; Ligand; Lipids; Identification; Gas analysis; Palmitic acid; Capture; Binding protein; Sample preparation; Extraction; Stearic acid; Dissociation; Sequencing; Electron capture detector; Collisional activation; Electron collision; Use; Ion cyclotron resonance spectrometry; Fatty acids; De novo; Variant; Gas chromatography; Mass spectrometry; Crystalline structure
• ISSN: 1618-2642; 1618-2650; 1618-2650
• DOI: 10.1007/s00216-010-3762-0

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. [graphic removed]