• RENTALA MADHUBALA

      Articles written in Journal of Biosciences

    • Deciphering the interaction of benzoxaborole inhibitor AN2690 with connective polypeptide 1 (CP1) editing domain of Leishmania donovani leucyl-tRNA synthetase

      SMRITI TANDON REETIKA MANHAS NEHA TIWARI MANOJ MUNDE RAMACHANDRAN VIJAYAN SAMUDRALA GOURINATH ROHINI MUTHUSWAMI RENTALA MADHUBALA

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      Leucyl-tRNA synthetases (LRS) catalyze the linkage of leucine with tRNALeu. A large insertion CP1 domain(Connective Polypeptide 1) in LRS is responsible for post-transfer editing of mis-charged aminoacyl-tRNAs.Here, we characterized the CP1 domain of Leishmania donovani, a protozoan parasite, and its role in editingactivity and interaction with broad spectrum anti-fungal, AN2690. The deletion mutant of LRS, devoid of CP1domain (LRS-CP1D) was constructed, followed by determination of its role in editing and aminoacylation.Binding of AN2690 and different amino acids with CP1 deletion mutant and full length LRS was evaluatedusing isothermal titration calorimetry (ITC) and molecular dynamics simulations. The recombinant LRS-CP1 Deltaprotein did not catalyze the aminoacylation and the editing reaction when compared to full-length LRS. Thus,indicating that CP1 domain was imperative for both aminoacylation and editing activities of LRS. Bindingstudies with different amino acids indicated selectivity of isoleucine by CP1 domain over other amino acids.These studies also indicated high affinity of AN2690 with the editing domain. Molecular docking studiesindicated that AN2690-CP1 domain complex was stabilized by hydrogen bonding and hydrophobic interactionsresulting in high binding affinity between the two. Our data suggests CP1 is crucial for the function of L.donovani LRS.

    • Rapid diagnosis of Leishmania infection with a portable loop-mediated isothermal amplification device

      MADHU PURI HARSIMRAN KAUR BRAR NIMISHA MITTAL EVANKA MADAN RAJESH SRINIVASAN KAPIL RAWAT SRIJA MOULIK MITALI CHATTERJEE SAI SIVA GORTHI ROHINI MUTHUSWAMI RENTALA MADHUBALA

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      L. donovani is an intracellular protozoan parasite, that causes visceral leishmaniasis (VL), and consequently,post-kala azar dermal leishmaniasis (PKDL). Diagnosis and treatment of leishmaniasis is crucial for decreasingits transmission. Various diagnostic techniques like microscopy, enzyme-linked immunosorbent assays(ELISA) and PCR-based methods are used to detect leishmaniasis infection. More recently, loop-mediatedisothermal amplification (LAMP) assay has emerged as an ideal diagnostic measure for leishmaniasis, primarilydue to its accuracy, speed and simplicity. However, point-of-care diagnosis is still not been tested withthe LAMP assay. We have developed a portable LAMP device for the monitoring of Leishmania infection. TheLAMP assay performed using our device can detect and amplify as little as 100 femtograms of L. donovaniDNA. In a preliminary study, we have shown that the device can also amplify L. donovani DNA present in VLand PKDL patient samples with high sensitivity (100%), specificity (98%) and accuracy (99%), and can beused both for diagnostic and prognostic analysis. To our knowledge, this is the first report to describe thedevelopment and application of a portable LAMP device which has the potential to evolve as a point-of-carediagnostic and prognostic tool for Leishmania infections in future.

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