title: Controlling gas-phase reactions for efficient charge reduction electrospray mass spectrometry of intact proteins.
abstract: Charge reduction electrospray mass spectrometry (CREMS) reduces the charge states of electrospray-generated ions, which concentrates the ions from a protein into fewer peaks spread over a larger m/z range, thereby increasing peak separation and decreasing spectral congestion. An optimized design for a CREMS source is described that provides an order-of-magnitude increase in sensitivity compared to previous designs and provides control over the extent of charge reduction. Either a corona discharge or an alpha-particle source was employed to generate anions that abstract protons from electrosprayed protein cations. These desired ion/ion proton transfer reactions predominated, but some oxidation and ion-attachment reactions also occurred, leading to new peaks or mass-shifted broader peaks while decreasing signal intensity. The species producing these deleterious side-reactions were identified, and conditions were found that prevented their formation. Spectrometer m/z biases were examined because of their effect upon the signal intensity of higher m/z charge-reduced protein ions. The utility of this atmospheric pressure CREMS was demonstrated using a cell lysate fraction from E. coli. The spectral simplification afforded by CREMS reveals more proteins than are observed without charge reduction.
authors.affiliation: Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, WI 53706, USA.
authors.forename: Brian L
authors.initials: BL
authors.lastname: Frey
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lin
authors.forename: Michael S
authors.initials: MS
authors.lastname: Westphall
authors.forename: Lloyd M
authors.initials: LM
authors.lastname: Smith
mesh_terms: D004563:Electrochemistry / Q000379:methods*; D005740:Gases / Q000032:analysis / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods*; D044367:Phase Transition; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D055672:Static Electricity
chemical_list: D005740:Gases; D011506:Proteins