Document Detail

Phosphoric acid as a matrix additive for MALDI MS analysis of phosphopeptides and phosphoproteins.
MedLine Citation:
PMID:  15373450     Owner:  NLM     Status:  MEDLINE    
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 alpha-casein and beta-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.
Sven Kjellström; Ole Nørregaard Jensen
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  76     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2004 Sep 
Date Detail:
Created Date:  2004-09-17     Completed Date:  2006-05-02     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5109-17     Citation Subset:  IM    
Copyright Information:
Copyright 2004 American Chemical Society
Protein Research Group, Department of Biochemistry & Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.
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MeSH Terms
Amino Acid Sequence
Bacterial Proteins / chemistry
Caseins / chemistry
Gentisates / chemistry
Ions / chemistry
Molecular Sequence Data
Peptide Fragments / analysis
Peptide Mapping / methods*
Phosphopeptides / analysis*
Phosphoproteins / chemistry*
Phosphoric Acids / chemistry*
Sensitivity and Specificity
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
Trypsin / chemistry
Reg. No./Substance:
0/Bacterial Proteins; 0/Caseins; 0/Gentisates; 0/Ions; 0/Peptide Fragments; 0/Phosphopeptides; 0/Phosphoproteins; 0/Phosphoric Acids; 490-79-9/2,5-dihydroxybenzoic acid; 7664-38-2/phosphoric acid; EC

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