| Autor: |
Fan QI; Program in Molecular and Cellular Cardiology, Department of Internal Medicine, Wayne State University School of Medicine, Detroit, MI 48201, USA., Vanderpool KM, O'Connor J, Marsh JD |
| Jazyk: |
angličtina |
| Zdroj: |
Molecular and cellular biochemistry [Mol Cell Biochem] 2003 Jan; Vol. 242 (1-2), pp. 3-10. |
| Abstrakt: |
To test the hypothesis that mutated beta2-subunits of the L-type calcium channel could serve as a decoy and interdict calcium channel trafficking and function, we engineered a beta2 subunit that contained the beta interaction domain for alpha1c 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 overexpressing mutated beta2 subunits in adult mammalian myocytes can markedly alter excitation-contraction coupling. This paradigm may offer new approaches to understanding and modulating EC coupling. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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