Summary
The recent introduction of electrospray ionization techniques that are suitable for peptides and whole proteins has allowed for the design of mass spectrometric protocols that provide accurate sequence information for proteins. The advantages gained by these approaches over traditional Edman Degradation sequencing include faster analysis and femtomole, sometimes attomole, sensitivity. The ability to efficiently identify proteins has allowed investigators to conduct studies on their differential expression or modification in response to various treatments or disease states. In this chapter, we discuss the use of electrospray tandem mass spectrometry, a technique whereby protein-derived peptides are subjected to fragmentation in the gas phase, revealing sequence information for the protein. This powerful technique has been instrumental for the study of proteins and markers associated with various disorders, including heart disease, cancer, and cystic fibrosis. We use the study of protein expression in cystic fibrosis as an example.
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Acknowledgements
The authors thank Junnan Chen and Samuel Shank for technical assistance. This work was funded by the Cystic Fibrosis Foundation. Dr. Ziady is an inventor on patents that cover the construction of a nonviral gene delivery system and holds equity in Copernicus Therapeutics Inc., a company that has licensed this technology. This presents no conflict for the work discussed in this chapter.
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Ziady, A.G., Kinter, M. (2009). Protein Sequencing with Tandem Mass Spectrometry. In: Foote, R., Lee, J. (eds) Micro and Nano Technologies in Bioanalysis. Methods in Molecular Biology™, vol 544. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-483-4_21
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DOI: https://doi.org/10.1007/978-1-59745-483-4_21
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