Mapping of bovine skeletal muscle proteins using two-dimensional gel electrophoresis and mass spectrometry.
The large individual variation in meat quality seen both within and between animals is not fully understood. Consequently, our long-term goal is to identify reliable proteins which control or determine bovine meat quality. Using a proteomic approach, bovine skeletal muscle samples were analyzed by two-dimensional gel electrophoresis (2-DE) using an immobilized pH 4-7 gradient in the first dimension and mass spectrometry. We first tested the reproducibility of the method. These experiments showed slightly greater intersample than intrasample variability. In order to evaluate the type of visualized proteins in 2-DE, we initiated the construction of a protein reference map of bovine Semitendinosus muscle. In total, 129 protein spots corresponding to 75 different gene products were identified. Of these proteins, the largest portion is involved in metabolism (25.5%), cell structure (17%), cell defense (16%) and contractile apparatus (14.5%). One quarter of the identified proteins are represented by two or several protein spots and multiple isoforms of troponin T are present. Peptide mass fingerprint results indicate that these isoforms are partly generated by alternative splicing. The data presented here are an important step for further proteome analyses on bovine muscle. This may lead to progress in understanding the mechanisms controlling postmortem muscle metabolism and meat quality.
Bouley
Julien
J
Unité de Recherches sur les Herbivores, Laboratoire Croissance et Métabolisme du Muscle, INRA, St-Genès-Champanelle, France.
Chambon
Christophe
C
Picard
Brigitte
B
D017398:Alternative Splicing; D000818:Animals; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry; D009124:Muscle Proteins / Q000302:isolation & purification*; D018482:Muscle, Skeletal / Q000737:chemistry*; D010449:Peptide Mapping / Q000379:methods*; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D020543:Proteome; D015203:Reproducibility of Results; D020107:Troponin T / Q000737:chemistry
D009124:Muscle Proteins; D020033:Protein Isoforms; D020543:Proteome; D020107:Troponin T