NMR spectroscopy of biomolecules
NMR spectroscopy of biomolecules
Dr. Jeetender Chugh obtained his PhD from Tata Institute of Fundamental Research (TIFR), Mumbai while working on protein assemblies using NMR spectroscopy, and method developments in NMR. He worked on two types of projects: (a) Structural characterization of the GTPase Effector Domain (GED) of dynamin, and (b) development of NMR methods to facilitate rapid resonance assignments in proteins.
Jeet did his post-doctoral research in the field of RNA-dynamics using NMR spectroscopy, where he studied the mechanism of A-site RNA populating an alternative conformation necessary for the proofreading mechanism in ribosomes. He also studied fluorinated peptides from biological perspective.
Structural Biology: RNA-protein interactions play a pivotal role in complex biological pathways. In literature, studies focussing on the structural characterisation of RNA-protein complexes are much less in number when compared to proteins alone or DNA-protein complexes thereby setting haze on the understanding of RNA-recognition mechanisms. My research group at IISER-Pune is primarily interested in gaining insights into the RNA-protein interactions using biophysical techniques wherein we measure atomic-level dynamics at multiple timescales using NMR spectroscopy. Why NMR? NMR spectroscopy can be used to measure dynamics at atomic resolution and to deduce structural, kinetics, and thermodynamics characteristics of many motional modes occurring at different timescales. NMR spectroscopy has broad sensitivity to motions spanning picosecond to second and longer timescales and can be used to characterize very subtle changes in conformation, including those involving minutely populated conformers that have exceptionally short lifetimes (in the order of nanoseconds). Last but not the least, NMR spectroscopy is a powerful approach for exploring how the dynamic structure landscape is modulated by cellular cues, and time-resolved methods can be used to follow these perturbations in real-time. Understanding the perturbations in complex biological pathways by using NMR- based metabolomics: The advantage NMR has in the field of metabolomics is that the quantitative accuracy is inherent in NMR due to the fact that integral of a signal is proportional to the number of protons of a given type, and it offers excellent reproducibility. Further, there is minimal sample preparation that is required and data can be acquired on cells, tissues, and any kind of body fluid. In this direction, we have used 1H-NMR based metabolomics in multiple conditions.
Yousf, S. & Chugh, J. (2021) Nuclear Magnetic Resonance Spectroscopy and Mass Spectrometry: Complementary Approaches to Analyze the Metabolome. Journal of Endocrinology and Reproduction, Vol:24, 21-30.
Datir, S. S. et al. (2020) Cold storage reveals distinct metabolic perturbations in processing and non-processing cultivars of potato (Solanum tuberosum L.). Scientific Reports, Vol: 10, 6268–13.
Rizvi, A. et al. (2019) Metabolomics studies to decipher stress responses in Mycobacterium smegmatispoint to a putative pathway of methylated amines biosynthesis. Journal of bacteriology. Vol: 201, 477–12.
Paithankar, H., Jadhav, P. V., Naglekar, A. S., Sharma, S. & Chugh, J. (2018) 1H, 13C and 15N resonance assignment of domain 1 of trans-activation response element (TAR) RNA binding protein isoform 1 (TRBP2) and its comparison with that of isoform 2 (TRBP1). Biomol Nmr Assigm Vol: 12, 189-194.
Chugh, J. (2014) Determining Transient Nucleic Acid Structures by NMR. in CHEMICAL BIOLOGY OF NUCLEIC ACIDS (Springer Berlin Heidelberg, 2014).