Dysferlin interacts with calsequestrin-1, myomesin-2 and dynein in human skeletal muscle
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- 作者：B¨¤rbara Flix ; Carolina de la Torre ; Juan Castillo ; Carme Casal ; Isabel Illa ; Eduard Gallardo
- 关键词：CALQ1 ; calsequestrin-1 ; Cav-3 ; caveolin-3 ; DHPR ; dihydropyridine receptor ; DMD ; Duchenne muscular dystrophy ; DYNC1LI2 ; cytoplasmic dynein 1 light intermediate chain 2 ; FLIM&ndash ; FRET ; fluorescence lifetime imaging&ndash ; fluorescence resonance energy tran
- 刊名：The International Journal of Biochemistry and Cell Biology
- 出版年：2013
- 出版时间：August, 2013
- 年：2013
- 卷：45
- 期：8
- 页码：1927-1938
- 全文大小：3209 K
文摘
Dysferlinopathies are a group of progressive muscular dystrophies characterized by mutations in the gene DYSF. These mutations cause scarcity or complete absence of dysferlin, a protein that is expressed in skeletal muscle and plays a role in membrane repair. Our objective was to unravel the proteins that constitute the dysferlin complex and their interaction within the complex using immunoprecipitation assays (IP), blue native gel electrophoresis (BN) in healthy adult skeletal muscle and healthy cultured myotubes, and fluorescence lifetime imaging-fluorescence resonance energy transfer (FLIM-FRET) analysis in healthy myotubes. The combination of immunoprecipitations and blue native electrophoresis allowed us to identify previously reported partners of dysferlin - such as caveolin-3, AHNAK, annexins, or Trim72/MG53 - and new interacting partners. Fluorescence lifetime imaging showed a direct interaction of dysferlin with Trim72/MG53, AHNAK, cytoplasmic dynein, myomesin-2 and calsequestrin-1, but not with caveolin-3 or dystrophin. In conclusion, although IP and BN are useful tools to identify the proteins in a complex, techniques such as fluorescence lifetime imaging analysis are needed to determine the direct and indirect interactions of these proteins within the complex. This knowledge may help us to better understand the roles of dysferlin in muscle tissue and identify new genes involved in muscular dystrophies in which the responsible gene is unknown.