Abstract
The pioneering work by Patrick H. O’Farrell established two-dimensional gel electrophoresis as one of the most important high-resolution protein separation techniques of modern biochemistry (Journal of Biological Chemistry 1975, 250, 4007–4021). The application of two-dimensional gel electrophoresis has played a key role in the systematic identification and detailed characterization of the protein constituents of skeletal muscles. Protein changes during myogenesis, muscle maturation, fibre type specification, physiological muscle adaptations and natural muscle aging were studied in depth by the original O’Farrell method or slightly modified gel electrophoretic techniques. Over the last 40 years, the combined usage of isoelectric focusing in the first dimension and sodium dodecyl sulfate polyacrylamide slab gel electrophoresis in the second dimension has been successfully employed in several hundred published studies on gel-based skeletal muscle biochemistry. This review focuses on normal and physiologically challenged skeletal muscle tissues and outlines key findings from mass spectrometry-based muscle proteomics, which was instrumental in the identification of several thousand individual protein isoforms following gel electrophoretic separation. These muscle-associated protein species belong to the diverse group of regulatory and contractile proteins of the acto-myosin apparatus that forms the sarcomere, cytoskeletal proteins, metabolic enzymes and transporters, signaling proteins, ion-handling proteins, molecular chaperones and extracellular matrix proteins.
Highlights
Efficient protein separation is a prerequisite for a variety of bioanalytical applications [1].Physicochemical parameters such as size, charge and solubility of individual polypeptides have been extensively exploited to develop sophisticated techniques for the isolation of specific protein species.Electro-focusing methods and one-dimensional gel electrophoresis (GE) are long established methods of protein biochemistry
In basic and applied myology, the application of the original O’Farrell method or slightly modified versions has resulted in the cataloging of several thousand distinct muscle protein isoforms and the identification of hundreds of fiber type-specific protein species
Gel-based proteomic studies have established a variety of protein changes in physiologically challenged skeletal muscles, including the effects of myogenesis, exercise, regeneration, hypoxia, prolonged disuse and natural aging
Summary
Efficient protein separation is a prerequisite for a variety of bioanalytical applications [1] Physicochemical parameters such as size, charge and solubility of individual polypeptides have been extensively exploited to develop sophisticated techniques for the isolation of specific protein species. This article focuses on the application of the 2D-GE method in basic myology and discusses the enormous scientific impact of this method on recent proteomic studies of normal and physiologically challenged skeletal muscle tissues. This includes the systematic cataloguing of the protein constituents of different contractile fiber types and the findings from surveys of proteome-wide changes during physiological adaptations. Sections give an overview of major proteomic studies that have employed 2D-GE methods and analyzed fiber type specification and protein changes during muscle development, fiber type transformation, exercise-induced adaptations, hypoxia-associated alterations, disuse-related muscular atrophy and skeletal muscle aging
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call