Battle for the EF-hands: Magnesium-calcium interference in calmodulin

A. Malmendal*, S. Linse, J. Evenäs, S. Forsén, T. Drakenberg

*Corresponding author

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Abstract

The ubiquitous Ca2+-regulatory protein calmodulin activates target enzymes as a response to submicromolar Ca2+ increases in a background of millimolar Mg2+. The potential influence of Mg2+/Ca2+ competition is especially intriguing for the N-terminal domain of the protein which possesses the sites with the lowest Ca2+ specificity. The interdependence of Ca2+ and Mg2+ binding in the N-terminal domain of calmodulin was therefore studied using 43Ca NMR, 1H-15N NMR, and fluorescent Ca2+ chelator techniques. The apparent affinity for Ca2+ was found to be significantly decreased at physiological Mg2+ levels. At Ca2+ concentrations of an activated cell the (Ca2+)2 state of the N-terminal domain is therefore only weakly populated, indicating that for this domain Ca2+ binding is intimately associated with binding of target molecules. The data are in good agreement with a two-site model in which each site can bind either Ca2+ or Mg2+. The Mg2+-Ca2+ binding interaction is slightly positively allosteric, resulting in a significantly populated (Mg2+)1(Ca2+)1 state. The Ca2+ off-rate from this state is determined to be at least one order of magnitude faster than from the (Ca2+)2 state. These two findings indicate that the (Mg2+)1(Ca2+)1 state is structurally and/or dynamically different from the (Ca2+)2 state. The 43Ca quadrupolar coupling constant and the 1H and 15N chemical shifts of the (Mg2+)1(Ca2+)1 state were calculated from titration data. The values of both parameters suggest that the (Mg2+)1(Ca2+)1 state has a conformation more similar to the 'closed' apo and (Mg2+)2 states than to the 'open' (Ca2+)2 state.
OriginalsprogEngelsk
TidsskriftBiochemistry
Vol/bind38
Udgave nummer36
Sider (fra-til)11844–11850
ISSN0006-2960
DOI
StatusUdgivet - 1999
Udgivet eksterntJa

Citer dette

Malmendal, A., Linse, S., Evenäs, J., Forsén, S., & Drakenberg, T. (1999). Battle for the EF-hands: Magnesium-calcium interference in calmodulin. Biochemistry, 38(36), 11844–11850. https://doi.org/10.1021/bi9909288