Page 69 - CSIR-IGIB Annual Report 2020-21

GTPases: complex regulator for Mtb H37Rv was done by INSTADOCK docking server in
survival which different GTPase inhibitors were used for
This project will help to understand the docking with FtsY protein. NAV2729 & ML141
significance of the GTP-binding family genes in showed highest binding affinity with FtsY. The
the survival and pathogenesis mechanism of activity was decreased at higher concentration
Mtb. Complete genome sequence of Mtb, of both inhibitors. These inhibitors were able to
suggested the presence of several homologs of inhibit bacterial growth at 200µM
bacterial GTP-binding proteins like ftsY, Era, concentration. Another study was done with
Obg, etc. Purified proteins will be checked for the PhoH2 which contains ATPase & GTPase
the presence of GTP binding and hydrolyzing activity which is optimized at 10mM Mg and
activity. Enzymatic assays, gene knockout and above Mg concentration decreases the activity.
Anti-sense approach would be used to assess ATP and ADP compete for binding on the active
the essentiality or functionality of genes site of PhoH2 and therefore ADP & GDP inhibits
belonging to GTP-binding family in the survival the ATPase & GTPase activity of PhoH2.
and pathogenesis strategy of Mtb. These results NAV2729 & ML141 decrease PhoH2 ATPase &
might confirm essentiality of these genes as GTPase activity. The antisense of PhoH2 affects
potential drug target. Achievements: FtsY and bacterial growth.
PhoH2 were purified, and its characterization Future Directions: The purified protein may
was done using GTPase assay and CD, ITC possess GTP binding or hydrolyzing activities,
respectively. The interaction between ligand therefore they might be essential for signaling
and FtsY was further evaluated by MD cascade driven by Mtb which may attribute to
simulation. Strong binding of ATP & GTP with its survival. Future experiments will involve
FtsY and weak binding of GDP and FtsY was disruption & anti-sense approach to evaluate
seen. Further to evaluate the essentiality of FtsY the essentiality of these genes in the survival
gene, it was cloned in pSTKT vector and and pathogenesis of Mtb. After disruption
antisense of FtsY was cloned in pSD5 vector. evaluation, we are planning for cell line studies
Magnesium (Mg) does not affect the growth to find the immunological parameters involved
much but increasing manganese concentration in stimulation by above mentioned genes.
decreases bacterial growth. Virtual screening

Publications
Analysis of predicted amino acid biosynthesis in Rv3344c in Mycobacterium tuberculosis H37Rv using
bioinformatics tools. Joshi K, Meena S and Meena* L.S., Biotechnol Appl Biochem, (2020) Mar; 67(2):213-223

Dual burden of tuberculosis (TB) and diabetes mellitus (DM) as the major risk factor for wide range of population.
Kumar A, Meena* L.S., Diabetes Obes Int J, (2020); 5(1); 000229 (1-8).

Multidirectional benefits of nanotechnology in the diagnosis, treatment and prevention of tuberculosis. Gupta M,
Shivangi and Meena* L.S., J Nanotechnol Nanomaterials, (2020); 1(2): 46-56.

Interconnection of Mycobacterium tuberculosis with host immune system. Kumar A, Shivangi, Agarwal P and
Meena* L.S., J Respir Dis Med, (2020); 2: 1-6.
Comprehensive analysis of GTP cyclohydrolase I activity in Mycobacterium tuberculosis H37Rv via Insilico studies.
Agarwal P and Meena* L.S. (2020) Biotechnol Appl Biochem, 2020 Jul 20.

Calcium ATPase signaling: A must include mechanism in the radar of therapeutics development against
tuberculosis. Shivangi and Meena* L.S. (2020) Biotechnol Appl Biochem, 2020 Sep 25.

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