title: Sheathless capillary electrophoresis-tandem mass spectrometry for top-down characterization of Pyrococcus furiosus proteins on a proteome scale.
abstract: Intact protein analysis via top-down mass spectrometry (MS) provides the unique capability of fully characterizing protein isoforms and combinatorial post-translational modifications (PTMs) compared to the bottom-up MS approach. Front-end protein separation poses a challenge for analyzing complex mixtures of intact proteins on a proteomic scale. Here we applied capillary electrophoresis (CE) through a sheathless capillary electrophoresis-electrospray ionization (CESI) interface coupled to an Orbitrap Elite mass spectrometer to profile the proteome from Pyrococcus furiosus. CESI-top-down MS analysis of Pyrococcus furiosus cell lysate identified 134 proteins and 291 proteoforms with a total sample consumption of 270 ng in 120 min of total analysis time. Truncations and various PTMs were detected, including acetylation, disulfide bonds, oxidation, glycosylation, and hypusine. This is the largest scale analysis of intact proteins by CE-top-down MS to date.
authors.affiliation: Department of Chemical Physiology, The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Han
authors.forename: Yueju
authors.initials: Y
authors.lastname: Wang
authors.forename: Aaron
authors.initials: A
authors.lastname: Aslanian
authors.forename: Marshall
authors.initials: M
authors.lastname: Bern
authors.forename: Mathieu
authors.initials: M
authors.lastname: Lavallée-Adam
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
mesh_terms: D019843:Archaeal Proteins / Q000032:analysis*; D019075:Electrophoresis, Capillary; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D020027:Pyrococcus furiosus / Q000737:chemistry*; D053719:Tandem Mass Spectrometry
chemical_list: D019843:Archaeal Proteins; D020543:Proteome