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Decoy calcium channel beta subunits modulate contractile function in myocytes

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Abstract

To test the hypothesis that mutated β2-subunits of the L-type calcium channel could serve as a decoy and interdict calcium channel trafficking and function, we engineered a β2 subunit that contained the beta interaction domain for α1c subunit interaction, but lacked N- and C-terminal domains that might be essential for sarcolemmal localization. An adenoviral vector was constructed containing the gene for the beta interaction domain (BID) fused to green fluorescence protein (GFP), using a vector containing only GFP as control. Freshly plated, dissociated adult rat myocytes were infected and expression and function were assessed at 60 h. Fluorescence microscopy confirmed GFP expression; immunoblot analysis confirmed dose-dependent GFP-BID expression. Mechanical properties of adult rat ventricular myocytes were evaluated using a video edge-detection system. Contractility analysis (optical/video, field stimulation) demonstrated that contracting cells decreased from 60 to 2%. Contractile amplitude (percent shortening) decreases significantly from 5.6 vs. 2.4% with no change in time to peak twitch. Recombinant adenovirus over-expressing mutated β2 subunits in adult mammalian myocytes can markedly alter excitation-contraction coupling. This paradigm may offer new approaches to understanding and modulating EC coupling.

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Authors and Affiliations

  1. Program in Molecular and Cellular Cardiology, Department of Internal Medicine, Wayne State University School of Medicine, Detroit, MI, USA

    Q. Ivy Fan, Kathleen M. Vanderpool & Jessica O'Connor

  2. Program in Molecular and Cellular Cardiology, Wayne State University School of Medicine, 421 East Canfield Avenue, Detroit, MI, 48201, USA

    James D. Marsh

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  1. Q. Ivy Fan
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  2. Kathleen M. Vanderpool
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  3. Jessica O'Connor
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  4. James D. Marsh
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Fan, Q.I., Vanderpool, K.M., O'Connor, J. et al. Decoy calcium channel beta subunits modulate contractile function in myocytes. Mol Cell Biochem 242, 3–10 (2003). https://doi.org/10.1023/A:1021128017974

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