Insertion of the full‐length calcium channel α1S subunit into triads of skeletal muscle in vitro

FEBS Letters

Flucher, Bernhard E¹  Kasielke, Nicole¹  Neuhuber, Birgit¹  Grabner, Manfred¹  Gerster, Uli¹ 

[1] Department of Physiology and Department of Biochemical Pharmacology, University of Innsbruck, Fritz-Pregl-Str. 3, A-6020 Innsbruck, Austria

关键词: Calcium channel;    Excitation-contraction coupling;    Immunofluorescence;    Skeletal muscle;    ER;    endoplasmic reticulum;    GFP;    green fluorescent protein;    α1S-GFP;    α1S with GFP at its C-terminus;    nt;    nucleotide number;    PCR;    polymerase chain reaction;    SR;    sarcoplasmic reticulum;    t-tubule;    transverse tubule;   

DOI  :  10.1016/S0014-5793(00)01583-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A full-length and a C-terminally truncated form of the calcium channel α_1S subunit can be isolated from skeletal muscle. Here we studied whether full-length α_1S is functionally incorporated into the skeletal muscle excitation-contraction coupling apparatus. A fusion protein of α_1S with the green fluorescent protein attached to its C-terminus (α_1S-GFP) or α_1S and GFP separately (α_1S+GFP) were expressed in dysgenic myotubes, which lack endogenous α_1S. Full-length α_1S-GFP was targeted into triad junctions and restored calcium currents and excitation-contraction coupling. GFP remained colocalized with α_1S, indicating that intact α_1S-GFP was inserted into triads and that the C-terminus remained associated with the excitation-contraction coupling apparatus.

【 授权许可】

Unknown