An optimized procedure for isolation of rodent and human skeletal muscle sarcoplasmic and myofibrillar proteins

Michael D Roberts; Kaelin C Young; Carlton D Fox; Christopher G Vann; Paul A Roberson; Shelby C Osburn; Johnathon H Moore; Petey W Mumford; Matthew A Romero; Darren T Beck; Cody T Haun; Veera L D Badisa; Benjamin M Mwashote; Victor Ibeanusi; Andreas N Kavazis

doi:10.14440/jbm.2020.307

An optimized procedure for isolation of rodent and human skeletal muscle sarcoplasmic and myofibrillar proteins

J Biol Methods. 2020 Feb 24;7(1):e127. doi: 10.14440/jbm.2020.307. eCollection 2020.

Authors

Michael D Roberts^{1

2}, Kaelin C Young^{1

2}, Carlton D Fox¹, Christopher G Vann¹, Paul A Roberson¹, Shelby C Osburn¹, Johnathon H Moore¹, Petey W Mumford¹, Matthew A Romero¹, Darren T Beck^{1

2}, Cody T Haun³, Veera L D Badisa⁴, Benjamin M Mwashote⁴, Victor Ibeanusi⁴, Andreas N Kavazis^{1

2}

Affiliations

¹ School of Kinesiology, Auburn University, Auburn, AL 36849, USA.
² Department of Cell Biology and Physiology, Edward Via College of Osteopathic Medicine Auburn Campus, Auburn, AL 36849, USA.
³ LaGrange College, LaGrange, GA 30240, USA.
⁴ School of the Environment, Florida A&M University, Tallahassee, FL 32307, USA.

Abstract

Several published protocols exist for isolating contractile or myofibrillar (MF) proteins from skeletal muscle, however, achieving complete resuspension of the myofibril pellet can be technically challenging. We performed several previously published MF isolation methods with the intent of determining which method was most suitable for MF protein isolation and solubilization. Here, we provide an optimized protocol to isolate sarcoplasmic and solubilized MF protein fractions from mammalian skeletal muscle suitable for several downstream assays.

Keywords: MF protein; actin; muscle; myosin; protein isolation.