L-type calcium channels and cytochrome b₅ reductase are components of protein complexes tightly associated with lipid rafts microdomains of the neuronal plasma membrane

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• 作者：Dorinda Marques-da-Silva ; Alej ; ro K. Samhan-Arias ; Teresa Tiago ; Carlos Gutierrez-Merino
• 关键词：L-type voltage-operated calcium channels ; Cytochrome b₅ reductase ; Lipid rafts ; Protein microdomains ; Resonance energy transfer ; Neurons
• 刊名：Journal of Proteomics
• 出版年：2010
• 出版时间：16 June 2010
• 年：2010
• 卷：73
• 期：8
• 页码：1502-1510
• 全文大小：598 K

文摘

The presence of cytosolic calcium microcompartments in neurons is well established. L-type voltage calcium channels play a leading role in the rise of cytosolic calcium in the neuronal soma and are sensitive to redox modulation. In a recent work [Samhan-Arias, A.K., García-Bereguiaín, M.A., Martín-Romero, F.J. and Gutiérrez-Merino, C. (2009) Mol. and Cell. Neurosci. 40, 14–26], we have shown that cytochrome b₅ reductase, whose deregulation leads to an overshot of superoxide anion production at the neuronal plasma membrane that triggers apoptosis in primary cultures of cerebellar granule neurons in culture, forms a large mesh of redox centres associated with lipid rafts in these neurons. In this work, we have implemented the use of fluorescent antibodies as reagents for quantitative Förster resonance energy transfer measurements and analysis using fluorescence microscopy images of cerebellar granule neurons in culture. The results of this study show that L-type voltage-operated calcium channels are also enriched in lipid rafts associated protein microdomains at a distance between 10 and 100 nm from cytochrome b₅ reductase. The methodological improvements done in this work can be also valuable for the study of proteins compartmentalization within other subcellular microdomains in any cell type in culture.