Calcium fingerprints induced by Calmodulin interactors in eukaryotic cells
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- 作者：Rania Dagher ; Christian Briè ; re ; Marie Fè ; ve ; Maria Zeniou ; Claire Pigault ; Christian Mazars ; Hervé ; Chneiweiss ; Raoul Ranjeva ; Marie-Claude Kilhoffer ; Jacques Haiech
- 关键词：Fluorescence polarization ; Fluorescence anisotropy ; SynCaM ; Fluorescent probe ; Calcium fingerprint ; Calmodulin ; Ca
- 刊名：Biochimica et Biophysica Acta (BBA)/Molecular Cell Research
- 出版年：2009
- 出版时间：June 2009
- 年：2009
- 卷：1793
- 期：6
- 页码：1068-1077
- 全文大小：1300 K
文摘
Calcium (Cap>2+ p>) is a ubiquitous second messenger which promotes cell responses through transient changes in intracellular concentrations. The prominent role of Cap>2+ p> in cell physiology is mediated by a whole set of proteins constituting a Cap>2+ p>-signalling toolkit involved in Cap>2+ p>-signal generation, deciphering and arrest. The different Cap>2+ p>-signalosomes deliver Cap>2+ p>-signals with spatial and temporal dynamics to control the function of specific cell types. Among the intracellular proteins involved in Cap>2+ p>-signal deciphering, calmodulin (CaM) plays a pivotal role in controlling Cap>2+ p>-homeostasis and downstream Cap>2+ p>-based signalling events. Due to its ubiquitous expression in eukaryotic cells and the variety of proteins it interacts with, CaM is central in Cap>2+ p>-signalling networks. For these reasons, it is expected that disrupting or modifying CaM interactions with its target proteins will affect Cap>2+ p>-homeostasis and cellular responses. The resulting calcium response will vary depending on which interactions between CaM and target proteins are altered by the molecules and on the specific Cap>2+ p>-toolkit expressed in a given cell, even in the resting state. In the present paper, the effect of six classical CaM interactors (W5, W7, W12, W13, bifonazole and calmidazolium) was studied on Cap>2+ p>-signalling in tumor initiating cells isolated from human glioblastoma (TG1) and tobacco cells (BY-2) using the fluorescent Cap>2+ p>-sensitive Indo-1 dye and aequorin, respectively. Various Cap>2+ p>-fingerprints were obtained depending both on the CaM interactor used and the cell type investigated. These data demonstrate that interaction between the antagonists and CaM results in a differential inhibition of CaM-dependent proteins involved in Cap>2+ p>-signal regulation. In addition, the distinct Cap>2+ p>-fingerprints in tobacco and human tumor initiating glioblastoma cells induced by a given CaM interactor highlight the specificity of the Cap>2+ p>-signalosome in eukaryotic cells.