![]() ![]() They separately calculated the pI values of those proteins from their amino acid sequences. In that study, the authors determined the focusing positions of 29 polypeptides of known amino acid sequence within a narrow range of immobilized pH gradients i.e., between pH 4.5 to 7.3 under denaturing conditions with 9.2 M and 9.8 M urea at 15 ☌ or 25 ☌, respectively. This web server calculates the pI values of proteins using pK values of amino acids as defined in, which were determined by examining polypeptide migration in an immobilized pH gradient (between pH 4.5 to 7.3) gel environment with 9.2 M and 9.8 M urea at 15 ☌ or 25 ☌. For this, we made use of the principle of the 2D gel electrophoresis, whereby, we computed the pI values using the “Compute pI/Mw tool – ExPASy” ( ) online webserver. In this study, we focused on the charge distribution (in terms of acidic and basic properties) of the translation factors throughout the domain Bacteria to comprehend the importance of the influence of the charge distribution of these factors on their accommodation on the ribosome and thus in their functions during this process of translation. However, in the broad aspect, which characteristics of the translation factors i.e., IF, EF and RF are necessary to be conserved for the accuracy of the universal process of protein synthesis among the different kinds of organisms need to be investigated. Based on several years of biochemical and structural biological studies worldwide, fairly detailed knowledge of the mechanisms of cellular protein synthesis is now known. Here, the accurate coordination of every participant protein factor is necessary to perform the process successfully. The protein factors that are involved in the successive events are initiation factors (IF), elongation factors (EF), release factors (RF), and ribosome recycling factors (RRF). With the help of specific protein factors and aminoacyl tRNAs, ribosomes carry out protein synthesis following the decoding of the genetic information from mRNA in successive events, namely, initiation, elongation, and termination (release and recycling). In Escherichia coli, 40% of the total cellular energy is utilized by the translation system. It is an energy-exhaustive cellular process. The translation is a complex universal biological process that takes place in a large macromolecular machine called ribosome in all living organisms. Furthermore, irrespective of the different shapes, sizes, and functions of the elongation and release factors, possession of the strictly acidic pI values of these translation factors all over the domain Bacteria indicates that the acidic nature of these factors is a necessary criterion, perhaps to interact into the partially enclosed rRNA rich inter-subunit space of the translating 70S ribosome. Acidic properties of these factors are independent of habitat, nature, and phylogeny of the bacterial species. Conclusionsįrom the results of our study, we conclude that among all the bacterial translation factors, elongation and release factors are more conserved in terms of their pI values in comparison to initiation and recycling factors. However, among all the translation factors, the translation initiation factor 2 (IF2) and ribosome recycling factor (RRF) showed variable pI values that are linked to the order of phylogeny. These acidic elongation factors and release factors contain higher frequencies of glutamic acids. We found that the translation initiation factors are mainly basic, whereas, elongation and release factors that interact with the inter-subunit space of the intact 70S ribosome during translation are strictly acidic across bacterial sp. Our study revealed the fact that it’s not all same, or all random, but there are distinct orders and the pI values of translation factors are translation event specific. Then we analyzed the output from various angles. In this regard, we asked the question about how similar are those translation factors to each other from a wide variety of bacteria? Hence, we did a thorough in silico study of the translation factors from 495 bacterial sp., and 4262 amino acid sequences by theoretically measuring their pI and MW values that are two determining factors for distinguishing individual proteins in 2D gel electrophoresis in experimental procedures. Protein synthesis is a cellular process that takes place through the successive translation events within the ribosome by the event-specific protein factors, namely, initiation, elongation, release, and recycling factors. ![]()
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