Advertisement

Isolation and Characterization of S100 Protein-Protein Complexes

  • Bence Kiss
  • Péter Ecsédi
  • Márton Simon
  • László NyitrayEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1929)

Abstract

S100 proteins are small, mostly dimeric, EF-hand Ca2+-binding proteins. Upon Ca2+ binding, a conformational change occurs resulting in the exposure of a shallow hydrophobic binding groove in each subunit. Interestingly, S100 proteins can interact with their partners in two ways: symmetrically, when the two partners identically bind into each groove, or asymmetrically, when only one partner binds to the S100 dimer occupying both binding pockets. Here we present a heterologous expression and purification protocol for all known human S100 proteins as well as for their partner peptides. Moreover, we provide a detailed description of three in vitro methods to determine the affinity, stoichiometry, and kinetics of S100 protein-protein interactions.

Key words

EF-hand Ca2+-binding Protein expression Protein purification Isothermal titration calorimetry (ITC) Fluorescence polarization (FP) Surface plasmon resonance (SPR) Fluorescence labelling S100 proteins 

Notes

Acknowledgment

This work was supported by the National Research, Development and Innovation Office (NKFIH) grants K119359 (to LN). MS was supported through the New National Excellence Program of the Hungarian Ministry of Human Capacities. We also thank the support of the MedInProt program of the Hungarian Academy of Sciences. Project no. FIEK_16-1-2016-0005 has been implemented with the support provided from the National Research, Development and Innovation Fund of Hungary, financed under the FIEK_16 funding scheme. This work was also supported by the National Development Agency Grant KMOP4.2.1/B-10-2011.

References

  1. 1.
    Boye K, Maelandsmo GM (2010) S100A4 and metastasis: a small actor playing many roles. Am J Pathol 176(2):528–535.  https://doi.org/10.2353/ajpath.2010.090526CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Bresnick AR, Weber DJ, Zimmer DB (2015) S100 proteins in cancer. Nat Rev Cancer 15(2):96–109.  https://doi.org/10.1038/nrc3893CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Yammani RR (2012) S100 proteins in cartilage: role in arthritis. Biochim Biophys Acta 1822(4):600–606.  https://doi.org/10.1016/j.bbadis.2012.01.006CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Donato R (2001) S100: a multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles. Int J Biochem Cell Biol 33(7):637–668CrossRefGoogle Scholar
  5. 5.
    Marenholz I, Heizmann CW, Fritz G (2004) S100 proteins in mouse and man: from evolution to function and pathology (including an update of the nomenclature). Biochem Biophys Res Commun 322(4):1111–1122.  https://doi.org/10.1016/j.bbrc.2004.07.096CrossRefPubMedGoogle Scholar
  6. 6.
    Ecsedi P, Kiss B, Gogl G, Radnai L, Buday L, Koprivanacz K, Liliom K, Leveles I, Vertessy B, Jeszenoi N, Hetenyi C, Schlosser G, Katona G, Nyitray L (2017) Regulation of the equilibrium between closed and open conformations of annexin A2 by N-terminal phosphorylation and S100A4-binding. Structure 25(8):1195–1207 e1195.  https://doi.org/10.1016/j.str.2017.06.001CrossRefPubMedGoogle Scholar
  7. 7.
    Gogl G, Alexa A, Kiss B, Katona G, Kovacs M, Bodor A, Remenyi A, Nyitray L (2016) Structural basis of ribosomal S6 kinase 1 (RSK1) inhibition by S100B protein: modulation of the extracellular signal-regulated kinase (ERK) signaling cascade in a calcium-dependent way. J Biol Chem 291(1):11–27.  https://doi.org/10.1074/jbc.M115.684928CrossRefPubMedGoogle Scholar
  8. 8.
    Kiss B, Duelli A, Radnai L, Kekesi KA, Katona G, Nyitray L (2012) Crystal structure of the S100A4-nonmuscle myosin IIA tail fragment complex reveals an asymmetric target binding mechanism. Proc Natl Acad Sci U S A 109(16):6048–6053.  https://doi.org/10.1073/pnas.1114732109CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Wang ZX (1995) An exact mathematical expression for describing competitive binding of two different ligands to a protein molecule. FEBS Lett 360(2):111–114.  https://doi.org/10.1016/0014-5793(95)00062-ECrossRefPubMedGoogle Scholar
  10. 10.
    Kiss B, Kalmar L, Nyitray L, Pal G (2016) Structural determinants governing S100A4-induced isoform-selective disassembly of nonmuscle myosin II filaments. FEBS J 283(11):2164–2180.  https://doi.org/10.1111/febs.13728CrossRefPubMedGoogle Scholar
  11. 11.
    Leclerc E, Fritz G, Weibel M, Heizmann CW, Galichet A (2007) S100B and S100A6 differentially modulate cell survival by interacting with distinct RAGE (receptor for advanced glycation end products) immunoglobulin domains. J Biol Chem 282(43):31317–31331.  https://doi.org/10.1074/jbc.M703951200CrossRefPubMedGoogle Scholar
  12. 12.
    Dattilo BM, Fritz G, Leclerc E, Kooi CW, Heizmann CW, Chazin WJ (2007) The extracellular region of the receptor for advanced glycation end products is composed of two independent structural units. Biochemistry 46(23):6957–6970.  https://doi.org/10.1021/bi7003735CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Ostendorp T, Leclerc E, Galichet A, Koch M, Demling N, Weigle B, Heizmann CW, Kroneck PM, Fritz G (2007) Structural and functional insights into RAGE activation by multimeric S100B. EMBO J 26(16):3868–3878.  https://doi.org/10.1038/sj.emboj.7601805CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Bence Kiss
    • 1
  • Péter Ecsédi
    • 1
  • Márton Simon
    • 1
  • László Nyitray
    • 1
    Email author
  1. 1.Department of BiochemistryELTE Eötvös Loránd UniversityBudapestHungary

Personalised recommendations