Time: To be announced
To be announced
Genome sequencing has led to the discovery of Type II Fatty acid synthesis pathway in Leishmania. An indispensable enzyme of the pathway is 4’ phosphopantetheinyl transferase (PPT) which catalyzes the transfer of 4’-phosphopantetheine arm from Coenzyme A to the conserved serine residue of the Acyl carrier protein. Phosphopantetheinyl transferase from other pathogens viz M. tuberculosis, P. aeroginosa have been shown to be important for the survival and pathogenecity of the microorganism. Since, Leishmania genome encodes a single PPT, it can act as a potential drug target and the understanding of the PPT as well ACP proteins may lead to the design of novel therapeutics against the deadly disease leishmaniasis.
Thus, the present study involves biophysical (Fluorescence, Circular Dichroism Spectroscopy) and biochemical characterization (Native PAGE, C18 reverse phase HPLC, Surface Plasmon Resonance) along with structure determination (X-ray crystallography and Nuclear Magnetic Resonance) of 4’ phosphopantetheinyl transferase (PPT) and its substrate Acyl carrier protein (ACP) of the type II fatty acid pathway of Leishmania. Followed by insights into the interaction interface of both the proteins. To speed up the search of a novel inhibitor the study also focuses on exploring the current state-of-the-art of drug repurposing strategies by screening small molecule chemical libraries as well as synthesized compounds against the Leishmania donovani promastigotes, axenic amastigotes and intramacrophagic stages of the parasite followed by calculation of the IC50 values and determining the cytotoxic effects of the molecules.
In future the hits obtained from whole cell based screening can be evaluated against the characterized fatty acid pathway proteins using in vitro enzymatic assays to determine the specific target of these molecules. We also aim to take forward these inhibitors to animal models of Leishmaniasis.