Summary
Protein noncovalent complexes compose most of the essential biological machines in the cell. The characterization of protein complexes and assemblies using mass spectrometry (MS) has significant advantages over many other biophysical methods because of the inherent sensitivity and resolution of MS. The applicability of MS coupled with electrospray ionization (ESI) for the measurement of large proteins and protein complexes has been furthered by the development of sensitive analyzers, such as the time-of-flight (TOF) analyzer. Moreover, sample preparation has a very important role for such studies, as it could significantly affect the results of mass spectrometry experiments. We discuss the experimental variables for the ESI-MS detection of noncovalent protein complexes by featuring two different protein systems: yeast enolase is a 93-kDa homodimeric complex, and α -synuclein is a small, 14-kDa protein implicated in the pathogenesis of Parkinson's disease and binds noncovalently to endogenous polyamines, such as spermine.
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Acknowledgments
We are grateful for the support for the UCLA Functional Pro-teomics Center provided by the W. M. Keck Foundation, and the funding of our research by the National Institutes of Health (RR 20004 to JAL).
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Yin, S., Loo, J.A. (2009). Mass Spectrometry Detection and Characterization of Noncovalent Protein Complexes. In: Lipton, M.S., Paša-Tolic, L. (eds) Mass Spectrometry of Proteins and Peptides. Methods In Molecular Biology, vol 492. Humana Press. https://doi.org/10.1007/978-1-59745-493-3_16
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DOI: https://doi.org/10.1007/978-1-59745-493-3_16
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