Abstract
The ability to purify cell organelles and protein complexes on a large scale, combined with advances in protein identification using mass spectrometry, has provided a wealth of information regarding protein localization and function. A major challenge in these studies has been the ability to identify bona fide organelle components from a background of co-purifying contaminants because none of the available biochemical purification protocols afford pure preparations. Since this situation is unlikely to change alternative strategies have been devised to meet this challenge by making use of the information inherent in the fractionation profile of organelles isolated by density gradient centrifugation. In this chapter we describe strategies based on protein correlation profiling and quantitative mass spectrometry to sort out likely candidates. The organelle inventories defined by these methods are suitable to guide future functional experiments.
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Acknowledgments
The research leading to these results has received funding from the European Commission’s 7th Framework Programme and grant agreements HEALTH-F4-2008-201648/PROSPECTS and HEALTH-F4-2007-200767/APO-SYS. JD was supported by the European Molecular Biology Organization. We thank all CEBI group members for helpful discussions and support.
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Dengjel, J., Jakobsen, L., Andersen, J.S. (2010). Organelle Proteomics by Label-Free and SILAC-Based Protein Correlation Profiling. In: Cutillas, P., Timms, J. (eds) LC-MS/MS in Proteomics. Methods in Molecular Biology, vol 658. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-780-8_15
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DOI: https://doi.org/10.1007/978-1-60761-780-8_15
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