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Targeted comparative proteomics by liquid chromatography/matrix-assisted laser desorption/ionization triple-quadrupole mass spectrometry.
Rapid Commun Mass Spectrom. 2006; 20(5):904-10.RC

Abstract

Here we report the first application of a matrix-assisted laser desorption/ionization (MALDI) triple-quadrupole mass spectrometer for targeted proteomics. Employing an amine-specific isotopic labelling approach, the technique was validated using five randomly selected bovine serum albumin peptides differentially labelled at known ratios. An indirect benefit of the isotopic labelling technique is a significant enhancement of the a1 ion in tandem mass (MS/MS) spectra of all peptides studied. Therefore, the a1 ion was selected as the fragment ion for multiple reaction monitoring (MRM) in all cases, eliminating tedious method development and optimization. Accurate quantification was achieved with an average relative standard deviation (RSD) of 5% (n = 5) and a detection limit of 14 amol. The technique was then applied to validate an important virulence biomarker of the fungal pathogen Candida albicans, which was not accurately quantified using global proteomics experiment employing two-dimensional liquid chromatography/electrospray ionization tandem mass spectrometry (2D-LC/ESI)-MS/MS. Using LC/MALDI-MRM analysis of five tryptic peptides, the protein PHR1 was found to be upregulated in the hyphal (pathogenic) form of C. albicans by a factor of 7.7 +/- 0.8.

Authors+Show Affiliations

Institute for Marine Biosciences, National Research Council of Canada, 1411 Oxford St., Halifax, Nova Scotia, Canada B3H 3Z1.No affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Validation Study

Language

eng

PubMed ID

16470697

Citation

Melanson, Jeremy E., et al. "Targeted Comparative Proteomics By Liquid Chromatography/matrix-assisted Laser Desorption/ionization Triple-quadrupole Mass Spectrometry." Rapid Communications in Mass Spectrometry : RCM, vol. 20, no. 5, 2006, pp. 904-10.
Melanson JE, Chisholm KA, Pinto DM. Targeted comparative proteomics by liquid chromatography/matrix-assisted laser desorption/ionization triple-quadrupole mass spectrometry. Rapid Commun Mass Spectrom. 2006;20(5):904-10.
Melanson, J. E., Chisholm, K. A., & Pinto, D. M. (2006). Targeted comparative proteomics by liquid chromatography/matrix-assisted laser desorption/ionization triple-quadrupole mass spectrometry. Rapid Communications in Mass Spectrometry : RCM, 20(5), 904-10.
Melanson JE, Chisholm KA, Pinto DM. Targeted Comparative Proteomics By Liquid Chromatography/matrix-assisted Laser Desorption/ionization Triple-quadrupole Mass Spectrometry. Rapid Commun Mass Spectrom. 2006;20(5):904-10. PubMed PMID: 16470697.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Targeted comparative proteomics by liquid chromatography/matrix-assisted laser desorption/ionization triple-quadrupole mass spectrometry. AU - Melanson,Jeremy E, AU - Chisholm,Kenneth A, AU - Pinto,Devanand M, PY - 2006/2/14/pubmed PY - 2006/4/7/medline PY - 2006/2/14/entrez SP - 904 EP - 10 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 20 IS - 5 N2 - Here we report the first application of a matrix-assisted laser desorption/ionization (MALDI) triple-quadrupole mass spectrometer for targeted proteomics. Employing an amine-specific isotopic labelling approach, the technique was validated using five randomly selected bovine serum albumin peptides differentially labelled at known ratios. An indirect benefit of the isotopic labelling technique is a significant enhancement of the a1 ion in tandem mass (MS/MS) spectra of all peptides studied. Therefore, the a1 ion was selected as the fragment ion for multiple reaction monitoring (MRM) in all cases, eliminating tedious method development and optimization. Accurate quantification was achieved with an average relative standard deviation (RSD) of 5% (n = 5) and a detection limit of 14 amol. The technique was then applied to validate an important virulence biomarker of the fungal pathogen Candida albicans, which was not accurately quantified using global proteomics experiment employing two-dimensional liquid chromatography/electrospray ionization tandem mass spectrometry (2D-LC/ESI)-MS/MS. Using LC/MALDI-MRM analysis of five tryptic peptides, the protein PHR1 was found to be upregulated in the hyphal (pathogenic) form of C. albicans by a factor of 7.7 +/- 0.8. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/16470697/Targeted_comparative_proteomics_by_liquid_chromatography/matrix_assisted_laser_desorption/ionization_triple_quadrupole_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -