Differential expression profiling of the proteomes and their mRNAs in porcine white and red skeletal muscles

Abstract

Skeletal muscle is an heterogeneous tissue with various biochemical and physical properties of several fiber types. In this study, we carried out the comparative study of protein expression patterns in white and red muscles using two-dimensional gel electrophoresis (2-DE). From more than 500 protein spots detected on each 2-DE gel, we screened five proteins that were differentially expressed between white and red muscles. Using peptide mass fingerprint and tandem mass spectrometry analysis these proteins were identified as myoglobin, two slow-twitch isoforms of myosin light chain and two small heat shock proteins (HSP20 and HSP27). The protein levels of myoglobin, myosin light chain and HSP20 were higher in red muscle, whereas HSP27 was higher in white muscle. In addition, genes of the identified proteins were cloned and their mRNAs were examined. Positive correlations between protein content and their mRNA levels were observed in white and red muscle. These results may provide us with valuable information to understand the different expression profiling between white and red muscle at the protein level.

Proteomic profiling of chronic low‐frequency stimulated fast muscle

“…The 2-D spot pattern of silver-stained gels was shown to be very comparable to published studies on the position of 2-D landmark species, such as contractile proteins [9][10][11][12][13][14][15][16][17][18], but the stimulated specimens differed considerably to the untreated control (Figs. 1A, C and E).…”

“…Due to its more heterogeneous nature, the proteomic profiling of skeletal muscle fibres is still in its infancy, but has already made sub-stantial contributions to our general understanding of basic and applied myology. Recent studies have successfully cataloged the skeletal muscle proteome from various animal species [9][10][11][12][13][14], determined global differences in protein expression between predominantly slow-versus fast-twitching fibres [15][16][17][18], and have identified novel marker proteins of muscle growth [19], myoblast differentiation [20], neonatal fibre necrosis [18], hypertrophy [21], muscular dystrophy [22][23][24][25][26], dysferlinopathy [27], immobilization-induced atrophy [28,29] and ageing-induced sarcopenia [30][31][32][33][34]. In analogy, based on the findings of an initial proteomic analysis of the fast-to-slow fibre transformation process using a conventional nonfluorescent method [35], this report describes the detailed DIGE analysis of the differential expression of the fast skeletal muscle proteome following chronic low-frequency stimulation.…”

Proteomic profiling of pathological and aged skeletal muscle fibres by peptide mass fingerprinting (Review)

“…9 was observed to a comparable degree in unstimulated versus transformed fibres. In agreement with the proteomic profiling of fast versus slow muscle populations (66)(67)(68)(69), the immunoblotting survey of biomarkers of the fast and slow muscle phenotype presented here demonstrates that the fast and slow isoforms of the sarcoplasmic reticulum Ca 2+ -ATPase and the terminal cisternae Ca 2+ -binding protein calsequestrin are reliable indicators of fibre type distribution. In addition, the expression of enzymes such as glyceraldehyde-phosphate dehydrogenase is a good marker for shifts from a glycolytic to a more oxidative metabolism (Fig.…”

Proteomic profiling of animal models mimicking skeletal muscle disorders

“…Recently, several studies on the proteomic profiling of animal models mimicking muscular dystrophy [41,[58][59][60][61] and sarcopenia [62][63][64][65] have been published. Since muscle fibres can be subdivided into different types of predominantly fast-or slow-twitching cells and because differentiated skeletal muscles remain highly plastic in their adaptive capabilities, the proteomics of fibre typing [66][67][68][69] and muscle transformation [70] has also been of considerable interest to cell biologists. Below we outline the results of these studies and describe the beginnings of skeletal muscle proteomics as a new biochemical field in basic and applied myology.…”