title: Separation of beta2-microglobulin conformers by high-field asymmetric waveform ion mobility spectrometry (FAIMS) coupled to electrospray ionisation mass spectrometry.
abstract: An investigation into the use of high-field asymmetric waveform ion mobility spectrometry (FAIMS) coupled to electrospray ionisation mass spectrometry (ESI-MS) for the differentiation of co-populated protein conformers has been conducted on the amyloidogenic protein beta(2)-microglobulin (beta(2)m). Accumulation of beta(2)m in vivo can result in the deposition of insoluble fibrils whose formation is thought to originate from partially folded protein conformers; hence, the folding properties of beta(2)m are of significant interest. We have analysed beta(2)m using ESI-FAIMS-MS under a range of pH conditions and have studied the effect of the ion mobility spectrometry parameters on the behaviour of the various protein conformers. The data show that different protein conformers can be detected and analysed by ESI-FAIMS-MS, the results being consistent with observations of pH denaturation obtained using complementary biophysical techniques. A variant of beta(2)m with different folding characteristics has been analysed for comparison, and the distinctions observed in the data sets for the two proteins are consistent with their folding behaviour. ESI-FAIMS-MS offers significant opportunities for the study of the conformational properties of proteins and thus may present valuable insights into the roles that different conformers play in diseases related to protein folding.
authors.affiliation: Astbury Centre for Structural Molecular Biology, School of Biochemistry & Microbiology, The University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Antoni J H
authors.initials: AJ
authors.lastname: Borysik
authors.forename: Paul
authors.initials: P
authors.lastname: Read
authors.forename: David R
authors.initials: DR
authors.lastname: Little
authors.forename: Robert H
authors.initials: RH
authors.lastname: Bateman
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
authors.forename: Alison E
authors.initials: AE
authors.lastname: Ashcroft
mesh_terms: D000595:Amino Acid Sequence; D045424:Complex Mixtures / Q000032:analysis / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017510:Protein Folding; D020033:Protein Isoforms / Q000032:analysis / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D001613:beta 2-Microglobulin / Q000032:analysis* / Q000737:chemistry* / Q000145:classification
chemical_list: D045424:Complex Mixtures; D020033:Protein Isoforms; D001613:beta 2-Microglobulin