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METHODS article
Front. Mol. Biosci.
Sec. Molecular Recognition
Volume 11 - 2024 |
doi: 10.3389/fmolb.2024.1398964
Application of Bio-Layer Interferometry for the Analysis of Ribosome-Protein Interactions
Provisionally accepted- Indian Institute of Technology Bombay, Mumbai, India
The ribosome, a ribonucleoprotein complex, performs the function of protein translation. While ribosomal RNA catalyzes polypeptide formation, several proteins assist the ribosome throughout the translation process. Studying the biochemical and kinetic properties of these proteins interacting with the ribosome is vital for elucidating their roles. Various techniques, such as zonal centrifugation, pull-down assays, dynamic light scattering (DLS), fluorescence polarization, and surface plasmon resonance (SPR) are employed for this purpose, each presenting unique advantages and limitations. We add to the repertoire of techniques by using Bio-Layer Interferometry (BLI) to examine interactions between the ribosome and translation factors. Our findings demonstrate that BLI can detect interactions of E. coli ribosomes with two proteins: E. coli initiation factor 2 (IF2) and P. falciparum translation enhancing factor (PTEF). A protein (Green Fluorescent Protein; GFP) known not to bind to E. coli ribosomes, shows no binding in the BLI assay. We show that BLI could be used to study the ribosome-protein interactions as it has key advantages like label-free procedures, ease of assay performance, and ribosome sample reuse. Our results highlight the comprehensive use of BLI in studying the ribosome-protein interactions, in addition to studying protein-protein and protein-ligand interactions.
Keywords: Ribosome-protein interaction, Bio-layer interferometry, binding affinity, Translation factors, ribosome
Received: 11 Mar 2024; Accepted: 08 Jul 2024.
Copyright: © 2024 Pandiarajan, Walunj, Banerjee, Rout, Srivastava, Patankar and Kaledhonkar. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Sujata B. Walunj, Indian Institute of Technology Bombay, Mumbai, India
Nirjhar Banerjee, Indian Institute of Technology Bombay, Mumbai, India
Swati Patankar, Indian Institute of Technology Bombay, Mumbai, India
Sandip Kaledhonkar, Indian Institute of Technology Bombay, Mumbai, India
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Sujata B. Walunj
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