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Phosphoric acid as a matrix additive for MALDI MS analysis of phosphopeptides and phosphoproteins

Kjellström, Sven LU and Jensen, Ole Nørregaard (2004) In Analytical Chemistry 76(17). p.5109-5117
Abstract
Phosphopeptides are often detected with low efficiency by MALDI MS analysis of peptide mixtures. In an effort to improve the phosphopeptide ion response in MALDI MS, we investigated the effects of adding low concentrations of organic and inorganic acids during peptide sample preparation in 2,5-dihydroxybenzoic acid (2,5-DHB) matrix. Phosphoric acid in combination with 2,5-DHB matrix significantly enhanced phosphopeptide ion signals in MALDI mass spectra of crude peptide mixtures derived from the phosphorylated proteins α-casein and β-casein. The beneficial effects of adding up to 1% phosphoric acid to 2,5-DHB were also observed in LC-MALDI-MS analysis of tryptic phosphopeptides of B. subtilis PrkC phosphoprotein. Finally, the mass... (More)
Phosphopeptides are often detected with low efficiency by MALDI MS analysis of peptide mixtures. In an effort to improve the phosphopeptide ion response in MALDI MS, we investigated the effects of adding low concentrations of organic and inorganic acids during peptide sample preparation in 2,5-dihydroxybenzoic acid (2,5-DHB) matrix. Phosphoric acid in combination with 2,5-DHB matrix significantly enhanced phosphopeptide ion signals in MALDI mass spectra of crude peptide mixtures derived from the phosphorylated proteins α-casein and β-casein. The beneficial effects of adding up to 1% phosphoric acid to 2,5-DHB were also observed in LC-MALDI-MS analysis of tryptic phosphopeptides of B. subtilis PrkC phosphoprotein. Finally, the mass resolution of MALDI mass spectra of intact proteins was significantly improved by using phosphoric acid in 2,5-DHB matrix. (Less)
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author
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
76
issue
17
pages
5109 - 5117
publisher
The American Chemical Society
external identifiers
  • scopus:4444236919
ISSN
1520-6882
DOI
10.1021/ac0400257
language
English
LU publication?
no
id
a2bf09de-c05d-475a-9373-261e7f4d18e2
date added to LUP
2016-12-09 20:52:02
date last changed
2017-10-22 05:23:34
@article{a2bf09de-c05d-475a-9373-261e7f4d18e2,
  abstract     = {Phosphopeptides are often detected with low efficiency by MALDI MS analysis of peptide mixtures. In an effort to improve the phosphopeptide ion response in MALDI MS, we investigated the effects of adding low concentrations of organic and inorganic acids during peptide sample preparation in 2,5-dihydroxybenzoic acid (2,5-DHB) matrix. Phosphoric acid in combination with 2,5-DHB matrix significantly enhanced phosphopeptide ion signals in MALDI mass spectra of crude peptide mixtures derived from the phosphorylated proteins α-casein and β-casein. The beneficial effects of adding up to 1% phosphoric acid to 2,5-DHB were also observed in LC-MALDI-MS analysis of tryptic phosphopeptides of B. subtilis PrkC phosphoprotein. Finally, the mass resolution of MALDI mass spectra of intact proteins was significantly improved by using phosphoric acid in 2,5-DHB matrix. },
  author       = {Kjellström, Sven and Jensen, Ole Nørregaard},
  issn         = {1520-6882},
  language     = {eng},
  number       = {17},
  pages        = {5109--5117},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Phosphoric acid as a matrix additive for MALDI MS analysis of phosphopeptides and phosphoproteins},
  url          = {http://dx.doi.org/10.1021/ac0400257},
  volume       = {76},
  year         = {2004},
}