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.
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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.
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Kiss, B., Ecsédi, P., Simon, M., Nyitray, L. (2019). Isolation and Characterization of S100 Protein-Protein Complexes. In: Heizmann, C. (eds) Calcium-Binding Proteins of the EF-Hand Superfamily. Methods in Molecular Biology, vol 1929. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9030-6_21
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DOI: https://doi.org/10.1007/978-1-4939-9030-6_21
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