Isolation and characterization of S100 protein-protein complexes

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

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (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

Fingerprint

S100 Proteins
Proteins
EF Hand Motifs
Peptides

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

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

Isolation and characterization of S100 protein-protein complexes. / Kiss, Bence; Ecsédi, Péter; Simon, Márton; Nyitray, L.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 325-338 (Methods in Molecular Biology; Vol. 1929).

Research output: Chapter in Book/Report/Conference proceedingChapter

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