Skeletal muscle is a plastic tissue with known gender dimorphism, especially at the metabolic level. A proteomic comparison of male and female murine biceps brachii was undertaken, resolving an average of 600 protein spots of MW 15–150 kDa and pI 5–8. Twenty-six unique full-length proteins spanning 11 KOG groups demonstrated statistically significant (p<0.05) abundance differences between genders; the majority of these proteins have metabolic functions. Identified glycolytic enzymes demonstrated decreased abundance in females, while abundance differences in identified oxidative phosphorylation enzymes were specific to the proteins rather than to the functional group as a whole. Certain cytoskeletal and stress proteins showed specific expression differences, and all three phosphorylation states of creatine kinase showed significant decreased abundance in females. Expression differences were significant but many were subtle (≤ 2-fold), and known hormonally-regulated proteins were not identified. We conclude that while gender dimorphism is present in non-exercised murine skeletal muscle, the proteome comparison of male and female biceps brachii in exercise-naive mice indicates subtle differences rather than a large or obviously hormonal dimorphism.
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The knowledge of concentration, modification and interaction of proteins is fundamental in determining the phenotype of living organisms. Plasma, the primary clinical specimen, contains numerous and diverse proteins. The functions of these proteins are as manifold as the diversity of the protein themselves. Many of them have been largely used for many years as biomarkers of diseases and indicators of the physiological functions. The study of plasma proteome promises to be a significant advance in various areas of biological and clinical research. Two-dimensional polyacrylamide gel electrophoresis is considered as a primary tool in separating thousand of plasma proteins. This approach enables comparing normal and diseased samples revealing differently expressed proteins. Other proteomic techniques suitable for plasma analysis such as protein microarrays are now either established or are still being improved. This article briefly reviews the application of two-dimensional electrophoresis and the current status of technical aspects for plasma proteome.
Frozen lung tissue sections from 2 healthy and 2 adenocarcinoma affected sheep were lysed in appropriate buffer. The two-dimensional (2D) electrophoresis of the protein lysates was performed. The resulting gels were visualised by silver and Coomassie Blue staining, then scanned and analysed using appropriate software. There was one spot present on the image obtained from the analysis of healthy tissue and no spot was found on cancer 2D gel image. The spot was excised and analysed using mass spectrometry. As a result, cytosolic NADP-isocitrate dehydrogenase was identified. In addition, several other protein spots of different intensity in neoplastic tissues, as compared with healthy ones, were found. The last finding reflects changes in protein expression in neoplastic and healthy tissues. These preliminary results can serve as the basis for more detailed investigations of the neoplastic tissue proteome, e.g.: isoelectric focusing in narrow pH range and analysis of correlation between tissue and serum protein profiles. The analysis of serum proteins from affected sheep can reveal markers of neoplastic process and help in preclinical diagnosis of ovine pulmonary adenocarcinoma.
We wstępie do artykułu przedstawiono stosunkowo nową dziedzinę badań – proteomikę, w obrębie której analizowane są: skład, budowa i funkcja białek oraz zachodzące pomiędzy nimi interakcje. W kolejnych rozdziałach opisano najważniejsze techniki analityczne proteomiki: elektroforezę dwukierunkową, metodę laserowej desorpcji/jonizacji próbki wspomaganej matrycą z analizatorem czasu przelotu (MALDI-TOF) oraz elektrorozpraszanie jonizacyjne (ESI, electrospray ionization). W celu przybliżenia zagadnień związanych z proteomiką w opracowaniu zaprezentowano przykłady analizy proteomu drobnoustrojów, w tym wybranych patogenów (Staphylococcus aureus, Vibrio cholerae, Bacillus subtilis, Mycobacterium avium, Borrelia ssp., Aspergillus flavus). Na podstawie wyników badań proteomu udokumentowano możliwość identyfikacji gatunku oraz badanie metabolizmu mikroorganizmu i interakcji gospodarz–patogen, a nawet możliwość różnicowania potencjału enzymatycznego mikroorganizmów utylizujących surowce ligninocelulozowe. W podsumowaniu zaznaczono potencjał analizy proteomicznej, przydatnej w wielostronnej charakterystyce drobnoustrojów.
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The establishment of technologies for high-throughput DNA sequencing (genomics), gene expression (transcriptomics), metabolite and ion analysis (metabolomics/ionomics) and protein analysis (proteomics) carries with it the challenge of processing and interpreting the accumulating data sets. Publicly accessible databases and newly development and adapted bioinformatic tools are employed to mine this data in order to filter relevant correlations and create models describing physiological states. These data allow the reconstruction of networks of interactions of the various cellular components as enzyme activities and complexes, gene expression, metabolite pools or pathway flux modes. Especially when merging information from transcriptomics, metabolomics and proteomics into consistent models, it will be possible to describe and predict the behaviour of biological systems, for example with respect to endogenous or environmental changes. However, to capture the interactions of network elements requires measurements under a variety of conditions to generate or refine existing models. The ultimate goal of systems biology is to understand the molecular principles governing plant responses and consistently explain plant physiology.
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