Thermodynamics of Calcium binding to the Calmodulin N-terminal domain to evaluate site-specific affinity constants and cooperativity
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| 024 | 7 | 0 | |a 10.1007/s00775-015-1275-1 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00775-015-1275-1 | ||
| 245 | 0 | 0 | |a Thermodynamics of Calcium binding to the Calmodulin N-terminal domain to evaluate site-specific affinity constants and cooperativity |h [Elektronische Daten] |c [Maria Beccia, Sandrine Sauge-Merle, David Lemaire, Nicolas Brémond, Romain Pardoux, Stéphanie Blangy, Philippe Guilbaud, Catherine Berthomieu] |
| 520 | 3 | |a Calmodulin (CaM) is an essential Ca(II)-dependent regulator of cell physiology. To understand its interaction with Ca(II) at a molecular level, it is essential to examine Ca(II) binding at each site of the protein, even if it is challenging to estimate the site-specific binding properties of the interdependent CaM-binding sites. In this study, we evaluated the site-specific Ca(II)-binding affinity of sites I and II of the N-terminal domain by combining site-directed mutagenesis and spectrofluorimetry. The mutations had very low impact on the protein structure and stability. We used these binding constants to evaluate the inter-site cooperativity energy and compared it with its lower limit value usually reported in the literature. We found that site I affinity for Ca(II) was 1.5 times that of site II and that cooperativity induced an approximately tenfold higher affinity for the second Ca(II)-binding event, as compared to the first one. We further showed that insertion of a tryptophan at position 7 of site II binding loop significantly increased site II affinity for Ca(II) and the intra-domain cooperativity. ΔH and ΔS parameters were studied by isothermal titration calorimetry for Ca(II) binding to site I, site II and to the entire N-terminal domain. They showed that calcium binding is mainly entropy driven for the first and second binding events. These findings provide molecular information on the structure-affinity relationship of the individual sites of the CaM N-terminal domain and new perspectives for the optimization of metal ion binding by mutating the EF-hand loops sequences. | |
| 540 | |a SBIC, 2015 | ||
| 690 | 7 | |a Calmodulin |2 nationallicence | |
| 690 | 7 | |a Calcium |2 nationallicence | |
| 690 | 7 | |a Microscopic binding constants |2 nationallicence | |
| 690 | 7 | |a Thermodynamics |2 nationallicence | |
| 690 | 7 | |a Cooperativity |2 nationallicence | |
| 700 | 1 | |a Beccia |D Maria |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | |
| 700 | 1 | |a Sauge-Merle |D Sandrine |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | |
| 700 | 1 | |a Lemaire |D David |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | |
| 700 | 1 | |a Brémond |D Nicolas |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | |
| 700 | 1 | |a Pardoux |D Romain |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | |
| 700 | 1 | |a Blangy |D Stéphanie |u Aix-Marseille Université, Architecture et Fonction des Macromolécules Biologiques, 13284, Marseille Cedex 09, France |4 aut | |
| 700 | 1 | |a Guilbaud |D Philippe |u CEA, DRCPC, SMCS, LILA, Bagnols Sur Cèze, France |4 aut | |
| 700 | 1 | |a Berthomieu |D Catherine |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | |
| 773 | 0 | |t JBIC Journal of Biological Inorganic Chemistry |d Springer Berlin Heidelberg |g 20/5(2015-07-01), 905-919 |x 0949-8257 |q 20:5<905 |1 2015 |2 20 |o 775 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00775-015-1275-1 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00775-015-1275-1 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Beccia |D Maria |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Sauge-Merle |D Sandrine |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Lemaire |D David |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Brémond |D Nicolas |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Pardoux |D Romain |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Blangy |D Stéphanie |u Aix-Marseille Université, Architecture et Fonction des Macromolécules Biologiques, 13284, Marseille Cedex 09, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Guilbaud |D Philippe |u CEA, DRCPC, SMCS, LILA, Bagnols Sur Cèze, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Berthomieu |D Catherine |u CEA, DSV, IBEB, Laboratoire des Interactions Protéine Métal, Saint-Paul-Lez-Durance, France |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t JBIC Journal of Biological Inorganic Chemistry |d Springer Berlin Heidelberg |g 20/5(2015-07-01), 905-919 |x 0949-8257 |q 20:5<905 |1 2015 |2 20 |o 775 | ||