title: Exploring the inhibitory activity of Withaferin-A against Pteridine reductase-1 of L. donovani.
abstract: Withaferin A is an abundant withanolide present in Withania somnifera leaves and to some extent in roots. It has been known for its profound anti-cancer properties, but its role in counteracting the Leishmania donovani infection has to be explored. Pteridine reductase 1 (PTR1) is involved in pteridine salvage and an important enzyme for the parasite growth, which could be targeted for the development of an efficient antileishmanial drug. We employed molecular docking studies to identify the binding mode of withaferin A with PTR1 in silico. We further cloned, expressed, and purified PTR1 of L. donovani and performed the enzyme kinetics using the Michaelis-Menten equation and enzyme inhibition studies with withaferin A by plotting the Lineweaver-Burk graph, which followed an uncompetitive mode of inhibition. We also showed the inhibition of the enzyme in the crude lysate of treated parasites. Thus, our study contributes towards understanding the mode of action of withaferin A against L. donovani parasite.
authors.affiliation: a Department of Animal Biology , University of Hyderabad , Hyderabad , Andhra Pradesh , India .
authors.forename: Sambamurthy
authors.initials: S
authors.lastname: Chandrasekaran
authors.affiliation: a Department of Animal Biology , University of Hyderabad , Hyderabad , Andhra Pradesh , India .
authors.forename: Jalaja
authors.initials: J
authors.lastname: Veronica
authors.affiliation: b Molecular Biology Unit , Institute of Medical Sciences, Banaras Hindu University , Varanasi , Uttar Pradesh , India , and.
authors.forename: Ravi Kumar
authors.initials: RK
authors.lastname: Gundampati
authors.affiliation: c Infectious Research Laboratory , Institute of Medical Sciences, Banaras Hindu University , Varanasi , Uttar Pradesh , India.
authors.forename: Shyam
authors.initials: S
authors.lastname: Sundar
authors.affiliation: a Department of Animal Biology , University of Hyderabad , Hyderabad , Andhra Pradesh , India .
authors.forename: Radheshyam
authors.initials: R
authors.lastname: Maurya
mesh_terms: D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D007893:Leishmania donovani / Q000201:enzymology*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors*; D054358:Withanolides / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D054358:Withanolides; D010088:Oxidoreductases; C085592:pteridine reductase; C009684:withaferin A
keywords: Autodock; Leishmania donovani; Pteridine reductase 1; Withaferin A; uncompetitive inhibition