title: Automated proteomics of E. coli via top-down electron-transfer dissociation mass spectrometry.
abstract: Electron-transfer dissociation (ETD) has recently been introduced as a fragmentation method for peptide and protein analysis. Unlike collisionally induced dissociation (CID), fragmentation by ETD occurs randomly along the peptide backbone. With the use of the sequences determined from the protein termini and the parent protein mass, intact proteins can be unambiguously identified. Because of the fast kinetics of these reactions, top-down proteomics can be performed using ETD in a linear ion trap mass spectrometer on a chromatographic time scale. Here we demonstrate the utility of ETD in high-throughput top-down proteomics using soluble extracts of E. coli. Development of a multidimensional fractionation platform, as well as a custom algorithm and scoring scheme specifically designed for this type of data, is described. The analysis resulted in the robust identification of 322 different protein forms representing 174 proteins, comprising one of the most comprehensive data sets assembled on intact proteins to date.
authors.affiliation: Mass Spectrometry Research Program, Research Triangle Institute, 3040 Cornwallis Road, Research Triangle Park, North Carolina 27709, USA.
authors.forename: Maureen K
authors.initials: MK
authors.lastname: Bunger
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Cargile
authors.forename: Anne
authors.initials: A
authors.lastname: Ngunjiri
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Bundy
authors.forename: James L
authors.initials: JL
authors.lastname: Stephenson
mesh_terms: D001331:Automation; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D040901:Proteomics / Q000379:methods*
chemical_list: D029968:Escherichia coli Proteins