title: Evaluation of an integrated strategy for proteomic profiling of skeletal muscle.
abstract: Proteomic analysis of skeletal muscle presents particular challenges when trying to identify valid biomarkers of phenotypic change in small biopsies from genetically diverse human subjects. Currently, two-dimensional (2-D) gel electrophoresis and mass spectrometry are the chosen analytical strategies but 2-D gels are not appropriate for analyzing proteins less than 11 kDa, they can suffer from problems of reproducibility and in routine use are not a viable high-throughput technique. We have evaluated an integrated proteomic strategy employing Ciphergen ProteinChip arrays, one-dimensional polyacrylamide gel electrophoresis and mass spectrometry. Protein fingerprints characteristic of fast and slow contracting muscles from normal and kyphoscoliosis (ky) mutant mice were obtained from Ciphergen protein arrays. Eight statistically validated protein biomarkers have so far been identified capable of discriminating fast from slow muscle. Five of these showed further differential expression in ky versus normal BDL soleus muscles. Several biomarkers have been formally identified, and were myosin light chain isoforms shown previously to be expressed differentially by fast versus slow skeletal muscles. This integrated experimental approach using a model mouse muscle system shows the potential of Ciphergen protein array technology for proteomic analysis of small proteins in small muscle samples and its applicability for phenotypic characterization of skeletal muscle in general.
authors.affiliation: Department of Basic Medical Sciences, St. George's Hospital Medical School, London, UK.
authors.forename: Marie-Catherine
authors.initials: MC
authors.lastname: Le Bihan
authors.forename: Edward
authors.initials: E
authors.lastname: Tarelli
authors.forename: Gary R
authors.initials: GR
authors.lastname: Coulton
mesh_terms: D000818:Animals; D015415:Biomarkers; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008817:Mice, Mutant Strains; D009124:Muscle Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D018482:Muscle, Skeletal / Q000737:chemistry*; D010449:Peptide Mapping; D040081:Protein Array Analysis; D011485:Protein Binding; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D015415:Biomarkers; D009124:Muscle Proteins; D020543:Proteome