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9 - Mass spectrometric techniques

Keith Wilson
Affiliation:
University of Hertfordshire
John Walker
Affiliation:
University of Hertfordshire
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Summary

INTRODUCTION

General

Mass spectrometry (MS) is an extremely valuable analytical technique in which the molecules in a test sample are converted to gaseous ions that are subsequently separated in a mass spectrometer according to their mass-to-charge (m/z) ratio and detected. The mass spectrum is a plot of the (relative) abundance of the ions at each m/z ratio. Note that it is the mass to charge ratios of ions (m/z) and not the actual mass that is measured. If for example, a biomolecule is ionised by the addition of one or more protons (H+ ions) the instrument measures the m/z after addition of 1 Da for each proton if the instrument is measuring positive ions or m/z minus 1 Da for each proton lost if measuring negative ions.

The development of two ionisation techniques, electrospray (ESI) and matrix-assisted laser desorption/ionisation (MALDI), has enabled the accurate mass determination of high-molecular-mass compounds as well as low-molecular-mass molecules and has revolutionised the applicability of mass spectrometry to almost any biological molecule. Applications include the new science of proteomics as well as in drug discovery. The latter includes combinatorial chemistry where a large number of similar molecules (combinatorial libraries) are produced and analysed to find the most effective compounds from a group of related organic chemicals.

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Publisher: Cambridge University Press
Print publication year: 2010

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References

Aebersold, R. and Mann, M. (2003). Mass spectrometry-based proteomics. Nature, 422, 198–207. (There are also a number of other, very informative proteomics reviews in this issue between pages 193 and 225.)CrossRefGoogle ScholarPubMed
Breitling, R., Pitt, A.R. and Barrett, M.P. (2006). Precision mapping of the metabolome. Trends in Biotechnology, 24, 543–548. (Review of the study of metabolic networks in complex mixtures using the high resolving power of FT-ICR and Orbitrap MS to discriminate between metabolites of near identical mass that differ perhaps by only 0.004 Da.)CrossRefGoogle ScholarPubMed
Glish, G. L. and Burinsky, D. J. (2008). Hybrid mass spectrometers for tandem mass spectrometry. Journal of the American Society for Mass Spectrometry, 19, 161–172. (Review of the development of hybrid mass spectrometers and a comparison of the particular applications of each type.)CrossRefGoogle ScholarPubMed
Hah, S. S. (2009). Recent advances in biomedical applications of accelerator mass spectrometry. Journal of Biomedical Science, 16, 54. (This is an Open Access article which reviews all aspects of this subject.)CrossRefGoogle ScholarPubMed
Han, X., Aslanian, A. and Yates, J.R. 3rd (2008). Mass spectrometry for proteomics. Current Opinion in Chemical Biology, 12, 1–8. (An excellent introductory review of the new instruments with a comparative table of their performance characteristics, such as mass resolution, mass accuracy, sensitivity, m/z range and main applications. Also covers quantitative proteomics and description of the latest jargon such as shotgun and top–down proteomics.)CrossRefGoogle ScholarPubMed
Nita-Lazar, A., Saito-Benz, H. and White, F. M. (2008). Quantitative phosphoproteomics by mass spectrometry: past, present, and future. Proteomics, 8, 4433–4443. (This reviews methods for identification of this important and widespread post-translational modification. This volume, no. 8 issue 21, 4367–4612 is a Special Issue on signal transduction proteomics which includes other informative articles on this subject.)CrossRefGoogle ScholarPubMed
The ExPASy (Expert Protein Analysis System) server of the Swiss Institute of Bioinformatics (SIB) contains a large suite of programs for the analysis of protein sequences, structures and proteomics as well as 2D PAGE analysis (2D gel documentation and 2D gel image analysis programmes). The ExPASy suite of programmes is at http://us.expasy.org/tools/
Glyocomod and GlycanMass are found at http://us.expasy.org/tools/glycomod/ and http://us.expasy.org/tools/glycomod/glycanmass.html respectively.
Deltamass is a database of protein post-translational modifications at http://www.abrf.org/index.cfm/dm.home which can be accessed to determine whether post-translational modifications are present. There are hyperlinks to references to the modifications.
There is also a prediction program ‘findmod’ for finding potential protein post-translational modifications in the ExPASy suite at http://expasy.org/tools/findmod/
Information and protocols for sample clean-up are found at http://www.millipore.com/catalogue/module/c5737 and http://www.nestgrp.com/protocols/protocol.shtml#massspec
Products for Phosphorylated Peptide and Protein Enrichment and Detection IMAC columns and chromatography at http://www.piercenet.com/files/phosphor.pdf and GelCode Phosphoprotein Staining Kit. http://www.piercenet.com/

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