COMPARISON OF EXTRACTION METHODS OF MUSCLE TISSUE PROTEINS
 
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Pol. J. Food Nutr. Sci. 2007;57(Special issue 4B):397–401
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ABSTRACT:
The aim of the study was to compare extraction methods of muscle tissue proteins, which could be applied to prepare samples for the assessment of meat proteins using two-dimensional electrophoresis (2-DE). Three methods were used: phenolic, with the use of the organic detergent CHAPS and drip forced by centrifugation. The assessment consisted in the determination which proteins are found in the extract and to what extent they correspond to proteins detected in the tissue. This evaluation was performed using polyacrylamide gel electrophoresis with SDS (PAGE-SDS) and urea. It was found that the highest number of proteins with the profile similar to that characterizing the tissue subjected to extraction was obtained as a result of the application of phenolic extraction, but only up to the time of extract purification. The process of extract purification caused considerable changes in the composition of the extract and limited the potential of its application in further analysis. Out of the other two methods the one based on the application of CHAPS turned out to be better. Analysis of centrifugal drip proteins offers the possibility of selective assessment of changes in cytoskeletal proteins with high molecular masses, which are normally absent. They appear as a result of processes occurring after slaughter. Results indicate that none of the applied extraction techniques makes it possible to obtain samples corresponding in their composition to muscle tissue proteins; however, the most similar results were recorded when using extraction with CHAPS. The selection of the method of extraction for analyses needs to take into consideration both the type of proteins to be investigated and the fact which method of extraction facilitates the presentation of their changes in the best possible way.