Abstract
Maturation of protein N-termini occurs in all kingdoms of life, with major protein modifications being proteolytic processing (e.g., removal of initiator methionines) and N-terminal acetylation. The functional consequences of these modifications are only known for a few substrates, and techniques to study such modifications have begun to emerge only recently. We here report on a method enabling targeted, mass spectrometry based analysis of protein N-termini from polyacrylamide gel-separated proteins. In our method, stable isotope incorporation by in-gel N-acetylation of free primary amines permits calculating the extent of in vivo N-terminal acetylation, proven to reveal crucial information with reference to N-terminal protein biology.
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Acknowledgments
P.V.D. is a Postdoctoral Fellow of the Research Foundation—Flanders (FWO-Vlaanderen). T.A. is supported by the Norwegian Research Council (Grant 197136) and the Norwegian Cancer Society. K.G. acknowledges support of research grants from the Fund for Scientific Research—Flanders (Belgium) (project numbers G.0042.07 and G.0440.10), the Concerted Research Actions (project BOF07/GOA/012) from the Ghent University and the Inter University Attraction Poles (IUAP06).
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Van Damme, P., Arnesen, T., Ruttens, B., Gevaert, K. (2013). In-Gel N-Acetylation for the Quantification of the Degree of Protein In Vivo N-Terminal Acetylation. In: Hake, S., Janzen, C. (eds) Protein Acetylation. Methods in Molecular Biology, vol 981. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-305-3_9
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DOI: https://doi.org/10.1007/978-1-62703-305-3_9
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-305-3
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