Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Dec 18;10(12):2743-52.
doi: 10.1021/acschembio.5b00616. Epub 2015 Oct 5.

Evaluation of Kinase Activity Profiling Using Chemical Proteomics


Evaluation of Kinase Activity Profiling Using Chemical Proteomics

Benjamin Ruprecht et al. ACS Chem Biol. .


Protein kinases are important mediators of intracellular signaling and are reversibly activated by phosphorylation. Immobilized kinase inhibitors can be used to enrich these often low-abundance proteins, to identify targets of kinase inhibitors, or to probe their selectivity. It has been suggested that the binding of kinases to affinity beads reflects a kinase's activation status, a concept that is under considerable debate. To assess the merits of the idea, we performed a series of experiments including quantitative phosphoproteomics and purification of kinases by single or mixed affinity matrices from signaling activated or resting cancer cells. The data show that mixed affinity beads largely bind kinases independent of their activation status, and experiments using individual immobilized kinase inhibitors show mixed results in terms of preference for binding the active or inactive conformation. Taken together, activity- or conformation-dependent binding to such affinity resins depends (i) on the kinase, (ii) on the affinity probe, and (iii) on the activation status of the lysate or cell. As a result, great caution should be exercised when inferring kinase activity from such binding data. The results also suggest that assaying kinase activity using binding data is restricted to a limited number of well-chosen cases.

Similar articles

See all similar articles

Cited by 10 articles

  • High-Throughput Assessment of Kinome-wide Activation States.
    Schmidlin T, Debets DO, van Gelder CAGH, Stecker KE, Rontogianni S, van den Eshof BL, Kemper K, Lips EH, van den Biggelaar M, Peeper DS, Heck AJR, Altelaar M. Schmidlin T, et al. Cell Syst. 2019 Oct 23;9(4):366-374.e5. doi: 10.1016/j.cels.2019.08.005. Epub 2019 Sep 11. Cell Syst. 2019. PMID: 31521607 Free PMC article.
  • The gut microbiota promotes hepatic fatty acid desaturation and elongation in mice.
    Kindt A, Liebisch G, Clavel T, Haller D, Hörmannsperger G, Yoon H, Kolmeder D, Sigruener A, Krautbauer S, Seeliger C, Ganzha A, Schweizer S, Morisset R, Strowig T, Daniel H, Helm D, Küster B, Krumsiek J, Ecker J. Kindt A, et al. Nat Commun. 2018 Sep 14;9(1):3760. doi: 10.1038/s41467-018-05767-4. Nat Commun. 2018. PMID: 30218046 Free PMC article.
  • Recent advances in methods to assess the activity of the kinome.
    Radu M, Chernoff J. Radu M, et al. F1000Res. 2017 Jun 26;6:1004. doi: 10.12688/f1000research.10962.1. eCollection 2017. F1000Res. 2017. PMID: 28713564 Free PMC article. Review.
  • Antimalarial efficacy of MMV390048, an inhibitor of Plasmodium phosphatidylinositol 4-kinase.
    Paquet T, Le Manach C, Cabrera DG, Younis Y, Henrich PP, Abraham TS, Lee MCS, Basak R, Ghidelli-Disse S, Lafuente-Monasterio MJ, Bantscheff M, Ruecker A, Blagborough AM, Zakutansky SE, Zeeman AM, White KL, Shackleford DM, Mannila J, Morizzi J, Scheurer C, Angulo-Barturen I, Martínez MS, Ferrer S, Sanz LM, Gamo FJ, Reader J, Botha M, Dechering KJ, Sauerwein RW, Tungtaeng A, Vanachayangkul P, Lim CS, Burrows J, Witty MJ, Marsh KC, Bodenreider C, Rochford R, Solapure SM, Jiménez-Díaz MB, Wittlin S, Charman SA, Donini C, Campo B, Birkholtz LM, Hanson KK, Drewes G, Kocken CHM, Delves MJ, Leroy D, Fidock DA, Waterson D, Street LJ, Chibale K. Paquet T, et al. Sci Transl Med. 2017 Apr 26;9(387):eaad9735. doi: 10.1126/scitranslmed.aad9735. Sci Transl Med. 2017. PMID: 28446690 Free PMC article.
  • Advances in mass spectrometry based strategies to study receptor tyrosine kinases.
    Vyse S, Desmond H, Huang PH. Vyse S, et al. IUCrJ. 2017 Feb 23;4(Pt 2):119-130. doi: 10.1107/S2052252516020546. eCollection 2017 Mar 1. IUCrJ. 2017. PMID: 28250950 Free PMC article. Review.
See all "Cited by" articles

LinkOut - more resources