Quantitative Analysis of Protein S-Acylation Site Dynamics Using Site-Specific Acyl-Biotin Exchange (ssABE)

  • Keith T. Woodley
  • Mark O. CollinsEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1977)


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.

Key words

Protein S-Acylation Palmitoylation Membrane Proteomic Mass spectrometry Quantitative 


  1. 1.
    Magee AI, Gutierrez L, McKay IA, Marshall CJ, Hall A (1987) Dynamic fatty acylation of p21N-ras. EMBO J 6(11):3353–3357CrossRefGoogle Scholar
  2. 2.
    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):125–134CrossRefGoogle Scholar
  3. 3.
    Blanc M, David F, Abrami L, Migliozzi D, Armand F, Burgi J et al (2015) SwissPalm: protein palmitoylation database. F1000Res 4:261CrossRefGoogle Scholar
  4. 4.
    Drisdel RC, Green WN (2004) Labeling and quantifying sites of protein palmitoylation. BioTechniques 36(2):276–285CrossRefGoogle Scholar
  5. 5.
    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):393–398CrossRefGoogle Scholar
  6. 6.
    Martin BR, Cravatt BF (2009) Large-scale profiling of protein palmitoylation in mammalian cells. Nat Methods 6(2):135–138CrossRefGoogle Scholar
  7. 7.
    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):246–258CrossRefGoogle Scholar
  8. 8.
    Collins MO, Woodley KT, Choudhary JS (2017) Global, site-specific analysis of neuronal protein S-acylation. Sci Rep 7(1):4683CrossRefGoogle Scholar
  9. 9.
    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):1367–1372CrossRefGoogle Scholar
  10. 10.
    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):4302–4307CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biomedical Science, Centre for Membrane Interactions and Dynamics (CMIAD)University of SheffieldSheffieldUK
  2. 2.Faculty of Science Mass Spectrometry CentreUniversity of SheffieldSheffieldUK

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