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First published online 25 April 2006
doi: 10.1242/jcs.02935
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Research Article |
1 Departments of Anaesthesia and Research, Basel University Hospital, Hebelstrasse 20, 4031 Basel, Switzerland
2 INSERM U607/CEA/UJF, Lab CCFP/DRDC, Rue des Martyrs 17, 38054, Grenoble, Cedex 9 France
3 Department of Physiology and Pharmacology, Gerontology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA
4 Department of Internal Medicine, Gerontology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA
5 Department of Experimental and Diagnostic Medicine, General Pathology Section, University of Ferrara, Via Borsari 46, 44100 Ferrara, Italy
* Author for correspondence (e-mail: zor{at}unife.it)
Accepted 14 February 2006
JP-45, an integral protein of the junctional face membrane of the skeletal muscle sarcoplasmic reticulum (SR), colocalizes with its Ca2+-release channel (the ryanodine receptor), and interacts with calsequestrin and the skeletal-muscle dihydropyridine receptor Cav1. We have identified the domains of JP-45 and the Cav1.1 involved in this interaction, and investigated the functional effect of JP-45. The cytoplasmic domain of JP-45, comprising residues 1-80, interacts with Cav1.1. JP-45 interacts with two distinct and functionally relevant domains of Cav1.1, the I-II loop and the C-terminal region. Interaction between JP-45 and the I-II loop occurs through the
-interacting domain in the I-II loop. ß1a, a Cav1 subunit, also interacts with the cytosolic domain of JP-45, and its presence drastically reduces the interaction between JP-45 and the I-II loop. The functional effect of JP-45 on Cav1.1 activity was assessed by investigating charge movement in differentiated C2C12 myotubes after overexpression or depletion of JP-45. Overexpression of JP-45 decreased peak charge-movement and shifted VQ1/2 to a more negative potential (-10 mV). JP-45 depletion decreased both the content of Cav1.1 and peak charge-movements. Our data demonstrate that JP-45 is an important protein for functional expression of voltage-dependent Ca2+ channels.
Key words: Voltage-dependent Ca2+ channel, JP-45, Sarcoplasmic reticulum, Excitation-contraction coupling
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