Abstract
Proteomic technologies have proven very effective for detection in meat products of biochemical changes, such as changes in heat resistant and species-specific proteins that could be relevant bio-markers. In the work presented in this report (for the period of 2013–2016), several tissue-specific proteins were detected in the samples of meat and specially developed meat products using proteomic technologies and identified as individual biomarkers in meat product control. The existence of a large number of different proteins resulted in the need to create information arrays — databases (or banks). Currently, there are a number of general and specialized databases that are available online to anyone interested. When studying protein proteomic profiles, many scientists stop at the stage of two-dimensional electrophoregrams sometimes even without ideas about the future prospects of using modern instruments and bioinformation resources to confirm or refute their hypotheses, and sometimes just to identify. This overview shows the chain of actions that allows going from profiling proteins in the gel to a specific interpretation of the results. Studies in this field have enabled formulating and significantly expanding the approaches to the identification and quantification of protein markers of quality, functionality and safe ty of meat raw material (detection of falsification) in the finished meat products. Based on the obtained data, the information was systematized using bioinformatics techniques with creation of the unique Atlas «Proteomic profiles of farm animal meat proteins.»
Highlights
The existence of a huge number of different proteins has led to the necessity of creating information arrays, i.e. databases, for entering all known information about them
The two-dimensional electrophoresis (2DE) by OFarrell with isoelectrofocusing in ampholine (IEFPAGE) or immobiline (IPG-PAGE) pH gradients is used as the main proteomic technology; the following protein detection is carried out by staining with Coomassie R-250 and silver nitrate [8, 12]
This can be presented schematically as follows (Fig. 1): Identification of protein fractions on two-dimensional electrophoregrams (2DE) is performed after tryptic proteolysis by MALDI-TOF MS and MS/MS mass-spectrometry using MALDI- TOF mass-spectrometer with UV-laser (336 nm) in the positive ion mode and a mass range of 500–1000 Da with their calibration according to the known trypsin autolysis peaks (Fig. 2)
Summary
The existence of a huge number of different proteins has led to the necessity of creating information arrays, i.e. databases, for entering all known information about them. The general databases contain information about all known proteins of living organisms, i.e., about the global proteome of all living things. Что, ввиду огромного структурного и функционального разнообразия, как белков, так и пептидов, для них до сих пор не создано строгой классификации. The decryption of the human genome, which represents an enormous matrix containing all information about humans and each cell of their organism, is time-consuming, torturous and even thankless task It is purely mechanical work, which is performed up to the bitter end and the result is a list of letters, a huge list, a book with letters and nothing more. Процесс нахождения новых мишеней для действия лекарств решается с помощью биоинформатики, причем объектом анализа является геном.
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