Abstract
Protein S-acylation (palmitoylation) is a reversible lipid modification that is increasingly recognized as an important regulator of protein function, including membrane association, trafficking, and subcellular localization. Most proteomic methods to study palmitoylation allow characterization of putative palmitoylated proteins but do not permit identification of individual sites of palmitoylation. We have recently adapted the Acyl-Biotin Exchange (ABE) method that is routinely used for palmitoyl-proteome characterization, to permit global S-acylation site analysis. This site-specific ABE (ssABE) protocol, when combined with SILAC-based quantification, allows both the large-scale identification of palmitoylation sites and quantitative profiling of palmitoylation site changes. This approach enables palmitoylation to be studied at a systems level comparable to other more intensively studied post-translational modifications.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Magee AI, Gutierrez L, McKay IA, Marshall CJ, Hall A (1987) Dynamic fatty acylation of p21N-ras. EMBO J 6(11):3353–3357
Topinka JR, Bredt DS (1998) N-terminal palmitoylation of PSD-95 regulates association with cell membranes and interaction with K+ channel Kv1.4. Neuron 20(1):125–134
Blanc M, David F, Abrami L, Migliozzi D, Armand F, Burgi J et al (2015) SwissPalm: protein palmitoylation database. F1000Res 4:261
Drisdel RC, Green WN (2004) Labeling and quantifying sites of protein palmitoylation. BioTechniques 36(2):276–285
Forrester MT, Hess DT, Thompson JW, Hultman R, Moseley MA, Stamler JS et al (2011) Site-specific analysis of protein S-acylation by resin-assisted capture. J Lipid Res 52(2):393–398
Martin BR, Cravatt BF (2009) Large-scale profiling of protein palmitoylation in mammalian cells. Nat Methods 6(2):135–138
Jones ML, Collins MO, Goulding D, Choudhary JS, Rayner JC (2012) Analysis of protein palmitoylation reveals a pervasive role in Plasmodium development and pathogenesis. Cell Host Microbe 12(2):246–258
Collins MO, Woodley KT, Choudhary JS (2017) Global, site-specific analysis of neuronal protein S-acylation. Sci Rep 7(1):4683
Cox J, Mann M (2008) MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol 26(12):1367–1372
Percher A, Ramakrishnan S, Thinon E, Yuan X, Yount JS, Hang HC (2016) Mass-tag labeling reveals site-specific and endogenous levels of protein S-fatty acylation. Proc Natl Acad Sci U S A 113(16):4302–4307
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Woodley, K.T., Collins, M.O. (2019). Quantitative Analysis of Protein S-Acylation Site Dynamics Using Site-Specific Acyl-Biotin Exchange (ssABE). In: Evans, C., Wright, P., Noirel, J. (eds) Mass Spectrometry of Proteins. Methods in Molecular Biology, vol 1977. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9232-4_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9232-4_6
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9231-7
Online ISBN: 978-1-4939-9232-4
eBook Packages: Springer Protocols