| DOI | Resolve DOI: https://doi.org/10.1002/rcm.2391 |
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| Author | Search for: Melanson, Jeremy E.1; Search for: Chisholm, Kenneth A.1; Search for: Pinto, Devanand M.1 |
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| Affiliation | - National Research Council of Canada. NRC Institute for Marine Biosciences
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| Format | Text, Article |
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| Subject | liquid chromatography; proteomics; electrospray ionization; mass spectrometry; matrix-assisted laser desorption ionization; methods; amino acid sequence; metabolism; Candida albicans; pathogens; cattle; albumin; bovine; virulence; markers |
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| Abstract | Here we report the first application of a matrix-assisted laser desorption/ionization (MALDI) triplequadrupole 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% (n¼5) 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 7.7W0.8. |
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| Publication date | 2006-02-08 |
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| Terms of use | - Copyright © 2006 John Wiley & Sons, Ltd.
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NRC number | NRCC 55833
1574 |
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| NPARC number | 3538457 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 3ae13d5c-91d3-4709-86f8-c6bbdfe5a42b |
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| Record created | 2009-03-01 |
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| Record modified | 2020-04-22 |
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