Prev. Article Next Article Table of Contents

Article

Phosphoric Acid as a Matrix Additive for MALDI MS Analysis of Phosphopeptides and Phosphoproteins

Sven Kjellström and Ole Nørregaard Jensen*

Protein Research Group, Department of Biochemistry & Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark

Anal. Chem., 2004, 76 (17), pp 5109–5117

DOI: 10.1021/ac0400257

Publication Date (Web): August 6, 2004

Corresponding author. Tel: +45 6550 2368. Fax: +45 6550 2467. URL: www.protein.sdu.dk. E-mail: jenseno@bmb.sdu.dk.

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.

View: Full Text HTML | Hi-Res PDF

Tools

SciFinder Links

History

Published In Issue September 01, 2004
Received for review February 13, 2004. Accepted June 9, 2004.

Recommend & Share

Related Content

Fe₃O₄/TiO₂ Core/Shell Nanoparticles as Affinity Probes for the Analysis of Phosphopeptides Using TiO₂ Surface-Assisted Laser Desorption/Ionization Mass SpectrometryAnalytical Chemistry
- Fe₃O₄/TiO₂ Core/Shell Nanoparticles as Affinity Probes for the Analysis of Phosphopeptides Using TiO₂ Surface-Assisted Laser Desorption/Ionization Mass Spectrometry
  Cheng-Tai Chen and Yu-Chie Chen
  Analytical Chemistry 2005 77 (18), pp 5912–5919
  Abstract: Columns packed with microsized titanium dioxide particles have been used effectively as precolumns for enriching phosphopeptides from complex mixtures. Nanosized titanium dioxide particles have a higher specific surface area and, hence, potentially higher ...
  Abstract | Full Text HTML | Hi-Res PDF
Selective Isolation at the Femtomole Level of Phosphopeptides from Proteolytic Digests Using 2D-NanoLC-ESI-MS/MS and Titanium Oxide PrecolumnsAnalytical Chemistry
- Selective Isolation at the Femtomole Level of Phosphopeptides from Proteolytic Digests Using 2D-NanoLC-ESI-MS/MS and Titanium Oxide Precolumns
  Martijn W. H. Pinkse, Pauliina M. Uitto, Martijn J. Hilhorst, Bert Ooms, and Albert J. R. Heck
  Analytical Chemistry 2004 76 (14), pp 3935–3943
  Abstract: Selective detection of phosphopeptides from proteolytic digests is a challenging and highly relevant task in many proteomics applications. Often phosphopeptides are present in small amounts and need selective isolation or enrichment before identification. ...
  Abstract | Full Text HTML | Hi-Res PDF
Enhanced Ionization of Phosphorylated Peptides during MALDI TOF Mass SpectrometryAnalytical Chemistry
- Enhanced Ionization of Phosphorylated Peptides during MALDI TOF Mass Spectrometry
  Xiaofeng Yang, Huaping Wu, Tomoyoshi Kobayashi, R. John Solaro, and Richard B. van Breemen
  Analytical Chemistry 2004 76 (5), pp 1532–1536
  Abstract: Although α-cyano-4-hydroxycinnamic acid functions as an excellent matrix for the analysis of most peptides using matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF) mass spectrometry, the ionization of phosphorylated peptides is usually ...
  Abstract | Full Text HTML | Hi-Res PDF

Article

Phosphoric Acid as a Matrix Additive for MALDI MS Analysis of Phosphopeptides and Phosphoproteins