title: Increasing protein charge state when using laser electrospray mass spectrometry.
abstract: Femtosecond (fs) laser vaporization is used to transfer cytochrome c, myoglobin, lysozyme, and ubiquitin from the condensed phase into an electrospray (ES) plume consisting of a mixture of a supercharging reagent, m-nitrobenzyl alcohol (m-NBA), and trifluoroacetic acid (TFA), acetic acid (AA), or formic acid (FA). Interaction of acid-sensitive proteins like cytochrome c and myoglobin with the highly charged ES droplets resulted in a shift to higher charge states in comparison with acid-stable proteins like lysozyme and ubiquitin. Laser electrospray mass spectrometry (LEMS) measurements showed an increase in both the average charge states (Zavg) and the charge state with maximum intensity (Zmode) for acid-sensitive proteins compared with conventional electrospray ionization mass spectrometry (ESI-MS) under equivalent solvent conditions. A marked increase in ion abundance of higher charge states was observed for LEMS in comparison with conventional electrospray for cytochrome c (ranging from 19+ to 21+ versus 13+ to 16+) and myoglobin (ranging from 19+ to 26+ versus 18+ to 21+) using an ES solution containing m-NBA and TFA. LEMS measurements as a function of electrospray flow rate yielded increasing charge states with decreasing flow rates for cytochrome c and myoglobin.
authors.affiliation: Department of Chemistry and Center for Advanced Photonics Research, Temple University, Philadelphia, PA, 19122, USA.
authors.forename: Santosh
authors.initials: S
authors.lastname: Karki
authors.forename: Paul M
authors.initials: PM
authors.lastname: Flanigan
authors.forename: Johnny J
authors.initials: JJ
authors.lastname: Perez
authors.forename: Jieutonne J
authors.initials: JJ
authors.lastname: Archer
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Levis
mesh_terms: D019342:Acetic Acid / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D001592:Benzyl Alcohols / Q000737:chemistry / Q000494:pharmacology; D002417:Cattle; D002645:Chickens; D045304:Cytochromes c / Q000737:chemistry*; D005561:Formates / Q000737:chemistry / Q000494:pharmacology; D006736:Horses; D007202:Indicators and Reagents / Q000737:chemistry / Q000494:pharmacology; D053844:Lasers, Solid-State; D009113:Muramidase / Q000737:chemistry*; D009211:Myoglobin / Q000737:chemistry*; D011489:Protein Denaturation / Q000187:drug effects; D055550:Protein Stability / Q000187:drug effects; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D014269:Trifluoroacetic Acid / Q000737:chemistry / Q000494:pharmacology; D025801:Ubiquitin / Q000737:chemistry*; D014835:Volatilization
chemical_list: D001592:Benzyl Alcohols; D005561:Formates; D007202:Indicators and Reagents; D009211:Myoglobin; D012997:Solvents; D025801:Ubiquitin; C030544:formic acid; D045304:Cytochromes c; D014269:Trifluoroacetic Acid; C421864:hen egg lysozyme; D009113:Muramidase; C046886:3-nitrobenzyl alcohol; D019342:Acetic Acid