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Volume 5, Issue 2 (Suppl) J Infect Dis Ther 2017
ISSN: 2332-0877, JIDT an open access journal Infection Congress 2017
May 11-12, 2017
May 11-12, 2017 Barcelona, Spain
4th International Congress on
Infectious Diseases
Characterization of phosphate transporter(s) and understanding their role in Leishmania donovani parasite
Ambak Kumar Rai, K J Sindhu, A K Kureel, S Saini, S Kumari and P Verma
Motilal Nehru National Institute of Technology, India
Statement of the Problem: Inorganic phosphate (Pi) is shown to be involved in excretion of methylglyoxal (MG) in the promastigote
form of Leishmania donovani parasite. Absence of Pi leads to its accumulation inside the parasite. Accumulation of MG is toxic to the
parasite and utilizes glyoxalase as well as excretory pathways for its detoxification. In addition, Pi is also reported to regulate activities
of ectoenzymes and energy metabolism (glucoseà pyruvate) etc. Thus, it cumulatively affects growth of the parasite. Therefore, the
transporters which allow the movement of Pi across the membrane can prove to be a crucial drug target.
Methodology & Theoretical Orientation: Bioinformatics analysis approach was applied to identify the phosphate transporters
in L. donovani. Phylogenetic analysis as well as secondary structure prediction was performed for its characterization. We tried
to understand the secondary structure of these two proteins and confirm modulation in their expression with the change in Pi
concentration outside. We also tried to detect the expression quantitatively in a log phase culture of promastigote and under various
physiological conditions. Moreover, their modes of action were also measured in presence of different inhibitors (LiF, CCCP).
Findings: We first characterized two phosphate transporters in Leishmania: H+ dependent myo-inositol transporter PHO84 and; Na+
dependent transporter PHO89 based on similar studies done previously on other lower organisms and trypanosomatids. We found
significantly higher expression of H+ dependent transporter (LdPHO84) as compared to Na+ dependent transporter (LdPHO89).
We also inhibited the Na+ as well as H+ gradient and then confirmed their involvement in the Pi transport across the membrane.
Conclusion & Significance: We thus functionally characterized two phosphate transporters of L .donovani and its regulation by
extracellular Pi for the first time. These computational results offer the possibility of identifying novel compounds which inhibit the
transport of Pi across the membrane.
Biography
Ambak Kumar Rai is an Assistant Professor in Department of Biotechnology at Motilal Nehru National Institute of Technology (MNNIT) Allahabad, India. He
has his expertise in “Understanding the molecular and biochemical aspects of drug resistance in Leishmania donovani”. A detailed approach to understand the
drug unresponsiveness offers new possibilities for therapeutic advancement. He is also inclined to understand the immuno-regulatory aspects of Leishmania
pathogenesis.
Ambak Kumar Rai et al., J Infect Dis Ther 2017, 5:2 (Suppl)