Protein NMR Spectroscopy:
Our lab specializes in the determination of atomic resolution structures of proteins in solution using state-of-the-art multidimensional
experiments of NMR spectroscopy. In addition to structure, we study the dynamics of protein molecules using NMR spectroscopy. We are also
carrying out NMR based screening of small molecule binders against protein targets. Currently, the focus of NMR structural work is as follows:
Bacterial peptidyl-tRNA hydrolases (Pth): Bacterial Pth proteins are validated targets for rational drug design of anti-microbial agents. In our lab, Pth from various mycobacterial and bacterial species are cloned, expressed, isotopically-labeled, and structurally characterized.
Eukaryotic Actin Depolymerization Factor (ADF)/Cofilin: Our lab is involved in NMR solution structure characterization of ADF/Cofilins from various eukaryotic species like Leishmania donovani, Toxoplasma gondii, C. elegans, D. melanogaster. We are also characterizing the proteins in which Cofilin fold is found, for ex. human plasma gelsolin G1, Glia Maturation Factor (GMF) etc.
Leishmania Rab proteins: We are involved in the challenging structural characterization of Leishmania Rab proteins. These proteins tend to precipitate at concentrations necessary for NMR spectroscopy, and over time. In order to prevent this, we are generating stable mutants of Rab proteins.
Biophysical Methods for characterization of proteins:
We study protein stability by Differential Scanning Calorimetry (DSC), Fluorescence spectroscopy, and CD spectroscopy. The protein-protein and protein-ligand interactions are characterized by Isothermal Titration Calorimetry.
Immunogenicity of Mycobacterial antigens:
Our lab is involved in characterization of secretary proteins from M. tuberculosis H37Rv. We have mainly targeted ESAT family proteins, ESAT-6, CFP-10, Rv3619, Rv3620, and MoaC1 (Rv3111). Lymphocyte proliferation and cytokine induction responses are studied for various antigens. Many of the antigens are structurally and biophysically characterized.
Protein crystallization and X-ray crystallography:
Our lab has achieved steady success in crystallization of many proteins like Pth, MoaC2, etc. Data is collected from various sources and crystal structures are determined after phasing through MR approach.
Characterization of peptides, antibiotics, and other molecules:
The characterization is done by using the standard 2D experiments of NMR spectroscopy, like DQF-COSY, TOCSY, NOESY, ROESY, HSQC, HMBC etc.
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