Tracing the footprints of Proteomics – To compare and study the techniques used in proteomics since the last decade.
Abstract: Proteomics is a study of the proteome of an organism. The last few decades have seen a rapid progress in the development of this field. This paper attempts to compare and contrast the way in which proteomics studies are performed today as opposed to those performed ten years ago and analyse its future implications.
The thrust of research while studying biology at a molecular level initially was focused specifically on the genomes of various organisms. As scientists discovered the intricacies of genes and their functionalities, the attention was soon drawn towards the end result of the central dogma of molecular biology, namely, the proteins, produced through translation of RNAs. Therefore, to study the proteins produced in an organism, referred to as the proteome, not just as products of a genome, but more importantly how they interact and bring about changes at the macro level, the field of proteomics has emerged. (1)
Proteins play a pivotal role in carrying out various functions in a body at the structural and dynamic levels. Proteins as enzymes and hormones regulate the vital metabolic processes and as structural components provide stability to the cellular components. The knowledge obtained through the study of these systems gives an insight into the overall functioning of the living organisms. In spite of having similar genetic blue prints, the protein expression in various organisms are regulated differently through diverse networks of protein-protein interactions. Hence, proteomics provides an understanding about these regulatory processes and establishes the differences and similarities between the evolutionary pathways of the organisms by grouping them under phylogentic trees. Further, drugs can be developed for specific diseases by designing structural analogues of proteins responsible for diseased conditions after elucidating their structures, which can then up or down regulate metabolic processes. Thus, the study of proteins plays an essential part of researches carried out in other related fields of study such as developmental and evolutionary biology and drug designing. (1)(2)
Since the invention of the 2-Dimentional Gel Electrophoresis in the 1970s, which is considered to be the stepping stone of modern day protein studies, scientists have been constantly striving to develop new and potent methods to study proteomics. Thus, this paper is an attempt to identify and compare these techniques which have been used and improved over the last decade. The popular and preferred procedure to study the proteome of an organism comprises of three major steps, isolation, separation on 2-D gel and analysis through a mass spectrometer. Most of the improvements revolve around this basic protocol.
2-D gel electrophoresis was one of the first methods which were used to analyse the proteome of an organism. In this technique, the protein is separated on the basis of its charge and size. The proteins are first separated on the basis of their different charges in the 1st dimension, following which they are separated on the 2nd dimension on the basis of their molecular weight. The gel or map provides a graphical representation of each protein after separation and hence they can be distinguished individually. However, the reproducibility of the results obtained through such an analysis has not been satisfactory. Till date there are constant efforts being made to improve the efficacy of this technique, such that a large number of proteins could be separated at the same time. The first 2-D separation which was carried out by using the electrophoresis buffer and starch gel, the improvements which followed gave rise to the foundation of modern day 2-D separation, which was combining two 1-d techniques involving separation on the basis of pH using isoelectric focusing (IEF) and using SDS-Page for...
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