Improvement of Shotgun Proteomics in the Negative Mode by Carbamylation of Peptides and Ultraviolet Photodissociation Mass Spectrometry | Zendy

Sylvester M. Greer | Zendy; Joe R. Can | Zendy; Jennifer S. Brodbelt | Zendy

Open Access

Improvement of Shotgun Proteomics in the Negative Mode by Carbamylation of Peptides and Ultraviolet Photodissociation Mass Spectrometry

Author(s) -

Sylvester M. Greer,

Joe R. Can,

Jennifer S. Brodbelt

Publication year - 2014

Publication title -

analytical chemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.117

H-Index - 332

eISSN - 1520-6882

pISSN - 0003-2700

DOI - 10.1021/ac5035314

Subject(s) - chemistry , mass spectrometry , chromatography , protein mass spectrometry , tandem mass spectrometry , electrospray ionization , derivatization , photodissociation , isobaric labeling , ultraviolet , bottom up proteomics , sample preparation in mass spectrometry , peptide , photochemistry , biochemistry , physics , quantum mechanics

Although acidic peptides compose a substantial portion of many proteomes, their less efficient ionization during positive polarity electrospray ionization (ESI) impedes their detection in bottom-up mass spectrometry workflows. We have implemented a derivatization strategy based on carbamylation which converts basic amine sites (Lys, N-termini) to less basic amides for enhanced analysis in the negative mode. Ultraviolet photodissociation (UVPD) is used to analyze the resulting peptide anions, as demonstrated for tryptic peptides from bovine serum albumin and Halobacterium salinarum in a high throughput liquid chromatography/tandem mass spectrometry (LC/MS/MS) mode. LC/UVPD-MS of a carbamylated H. salinarum digest resulted in 45% more identified peptides and 25% more proteins compared to the unmodified digest analyzed in the negative mode.

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