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Divers Models of Divalent Cation Interaction to Calcium-Binding Proteins: Techniques and Anthology

  • Jos A. CoxEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 963)

Abstract

Intracellular Ca2+-binding proteins (CaBPs) are sensors of the calcium signal and several of them even shape the signal. Most of them are equipped with at least two EF-hand motifs designed to bind Ca2+. Their affinities are very variable, can display cooperative effects, and can be modulated by physiological Mg2+ concentrations. These binding phenomena are monitored by four major techniques: equilibrium dialysis, fluorimetry with fluorescent Ca2+ indicators, flow dialysis, and isothermal titration calorimetry. In the last quarter of the twentieth century reports on the ion-binding characteristics of several abundant wild-type CaBPs were published. With the advent of recombinant CaBPs it became possible to determine these properties on previously inaccessible proteins. Here I report on studies by our group carried out in the last decade on eight families of recombinant CaBPs, their mutants, or truncated domains. Moreover this chapter deals with the currently used methods for quantifying the binding of Ca2+ and Mg2+ to CaBPs.

Key words

Equilibrium dialysis Fluorescent Ca2+ indicators Flow dialysis Isothermal titration calorimetry Penta-EF-hand proteins S100 proteins Invertebrate-specific CaBPs Centrins NOX5 

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Department of BiochemistryUniversity of GenevaGenevaSwitzerland

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