title: Separation and identification of rat skeletal muscle proteins using two-dimensional gel electrophoresis and mass spectrometry.
abstract: Skeletal muscle plays a major role in whole body protein metabolism, and changes in the rates of synthesis and degradation of proteins are likely to lead to characteristic changes in the amounts of different proteins in muscle under various physiological and pathological conditions. This paper demonstrates the feasibility of a proteomic approach to analyzing the protein composition of skeletal muscle. We report here the initial establishment of two-dimensional gel electrophoresis (2-D PAGE) reference maps for mixed skeletal muscle taken from the abdominal wall of a normal adult rat. We used immobilized pH gradients of 3-10 (non-linear) and 4-7 (linear), and matrix assisted laser desorption/ionization--time of flight mass spectrometry for protein identification by peptide mass fingerprinting. More than 600 protein spots were detected on each gel, of which 100 were excised and characterized. In-gel digestion followed by peptide mass fingerprinting enabled tentative identification of 74 of these, which included a wide range of myofibrillary and sarcoplasmic proteins. This database should provide the nucleus of a valuable resource for investigation of the biochemical basis of skeletal muscle pathologies in general and such specific disorders as alcoholic myopathy and injury.
authors.affiliation: Heart Science Centre, Imperial College School of Medicine, Harefield, Middlesex, UK. jun.yan@eu.apbiotech.com
authors.forename: J X
authors.initials: JX
authors.lastname: Yan
authors.forename: R A
authors.initials: RA
authors.lastname: Harry
authors.forename: R
authors.initials: R
authors.lastname: Wait
authors.forename: S Y
authors.initials: SY
authors.lastname: Welson
authors.forename: P W
authors.initials: PW
authors.lastname: Emery
authors.forename: V R
authors.initials: VR
authors.lastname: Preedy
authors.forename: M J
authors.initials: MJ
authors.lastname: Dunn
mesh_terms: D000818:Animals; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D009124:Muscle Proteins / Q000302:isolation & purification*; D018482:Muscle, Skeletal / Q000737:chemistry*; D010449:Peptide Mapping; D020543:Proteome; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D009124:Muscle Proteins; D020543:Proteome