Purification of sarcoplasmic reticulum vesicles from horse gluteal muscle.

ABSTRACT: We have analyzed protein expression and enzyme activity of the sarcoplasmic reticulum Ca²⁺-transporting ATPase (SERCA) in horse gluteal muscle. Horses exhibit a high incidence of recurrent exertional rhabdomyolysis, with myosolic Ca²⁺ proposed, but yet to be established, as the underlying cause. To better assess Ca²⁺ regulatory mechanisms, we developed an improved protocol for isolating sarcoplasmic reticulum (SR) vesicles from horse skeletal muscle, based on mechanical homogenization and optimized parameters for differential centrifugation. Immunoblotting identified the peak subcellular fraction containing the SERCA1 protein (fast-twitch isoform). Gel analysis using the Stains-all dye demonstrated that calsequestrin (CASQ) and phospholipids are highly enri

SUBMITTER: Autry JM 

PROVIDER: S-EPMC7745508 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature