Isolation and characterization of S100 protein-protein complexes

Bence Kiss, Péter Ecsédi, Márton Simon, L. Nyitray

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Citations (Scopus)

Abstract

S100 proteins are small, mostly dimeric, EF-hand Ca 2+ -binding proteins. Upon Ca 2+ 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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages325-338
Number of pages14
DOIs
Publication statusPublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1929
ISSN (Print)1064-3745

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Kiss, B., Ecsédi, P., Simon, M., & Nyitray, L. (2019). Isolation and characterization of S100 protein-protein complexes. In Methods in Molecular Biology (pp. 325-338). (Methods in Molecular Biology; Vol. 1929). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9030-6_21