Calcium Signalling pp 59-63 | Cite as
MCU Regulation in Lipid Bilayer and Electrophysiological Recording
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Abstract
The mitochondrial calcium uniporter (MCU) and the mitochondrial calcium uniporter dominant negative b- subunit (MCUb) are pore-forming components of the uniporter complex. We expressed these MCU subunits in cell-free transcription/translation systems, and we studied them, at the single molecule level, using the electrophysiological technique of planar lipid bilayer.
We showed that MCU gives rise to single-channel Ca2+ currents. In contrast, MCUb alone does not display calcium-permeable channel activity, while the co-expression of MCUb:MCU drastically alters the calcium permeation mediated by MCU subunit.
Key words
Planar lipid bilayer Ion channel Calcium signalingReferences
- 1.Vasington FD, Murphy JV (1962) Ca ion uptake by rat kidney mitochondria and its dependence on respiration and phosphorylation. J Biol Chem 237:2670–2677PubMedPubMedCentralGoogle Scholar
- 2.Rizzuto R, Simpson AW, Brini M, Pozzan T (1992) Rapid changes of mitochondrial Ca2+ revealed by specifically targeted recombinant aequorin. Nature. 1992;358(6384):325–7. Erratum in. Nature 360(6406):768CrossRefGoogle Scholar
- 3.Baughman JM, Perocchi F, Girgis HS et al (2011) Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature 476:341–345CrossRefGoogle Scholar
- 4.De Stefani D, Raffaello A, Teardo E et al (2011) A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature 476:336–340CrossRefGoogle Scholar
- 5.Raffaello A, De Stefani D, Sabbadin D et al (2013) The mitochondrial calcium uniporter is a multimer that can include a dominant-negative pore-forming subunit. EMBO J 32:2362–2376CrossRefGoogle Scholar
- 6.Chaudhuri D, Sancak Y, Mootha VK, Clapham DE. (2013) MCU encodes the pore conducting mitochondrial calcium currents. Elife. 4;2:e00704.Google Scholar
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