Qualification: 
MSc, BSc
divyanedungadi@am.amrita.edu

Ms Divya Nedungadi P after completing her M.Sc in Biotechnology from Amrita School of Biotechnology in 2012 joined us as a PhD Scholar in the Cancer Biology Lab. She is currently working in a Department of Science and Technology (DST) funded project titled “Paraptosis- A newer approach to target cancer” under Dr Nandita Mishra, Assistant Professor.

ALMA MATER

  • MSc. Biotechnology, Amrita Vishwa Vidyapeetham (2012)
  • BSc. Biotechnology, PSG College of Arts & Science, Coimbatore (2010)

AWARDS / HONOR

  • Award of Excellence from Amrita Vishwa Vidyapeetham,2013
  • Qualified ICMR-Junior Research Fellowship, 2012
  • Qualified GATE exam, 2012

Research experience

  • “Role of calpain and caspase-3 in regulating the mitochondrial fusion protein,Opa-1 during apoptosis”, Rajiv Gandhi Centre for Biotechnology, Kerala under Dr TR Santhoshkumar (Jan 2012-May 2012).
  • Protein purification at MIMS Research Foundation, Kerala under Dr R.V. Thampan (June 2011).
  • Recombinant DNA Technology and PCR, Shreedhar Bhatt Laboratory, Bangalore (May 2009)
     

Publications

Publication Type: Conference Proceedings

Year of Publication Title

2020

Anupama Binoy, Divya Nedungadi, Damu Sunilkumar, Dr. Bipin G. Nair, and Dr. Nandita Mishra, “Role of AIF in Plumbagin induced Paraptosis-A caspase independent cell death in MDA-MB-231 cancer cells”, 39th Annual Conference of Indian Association for Cancer Research (IACR-2020): “Leading the Fight against Cancer”, Rajiv Gandhi Centre for Biotechnology (RGCB), Trivandrum, Kerala, India, February 5-7. 2020.

2017

Divya Nedungadi, Megha Prasad, Anupama Binoy, Patra, G., Silva, J. D., Chatterjee, M., Jacob, A., Babu, A. S., Alexin, R., Dr. Bipin G. Nair, Dr. Sanjay Pal, and Dr. Nandita Mishra, “Anthelmintic properties of ginger compounds”, 27th Swadeshi Science Congress, November 7-9, 2017. Kerala, India, 2017.

2013

Dr. Nandita Mishra, Dr. Sanjay Pal, Nair, R. R., Krishna, A., Akhila Ajith, V, A., KS, D., Divya Nedungadi, and P, C., “Expression and refolding of recombinant staphylococcal amidases in E. coli”, Proceedings of International Conference on Biotechnology for Innovative Applications (Amrita BioQuest 2013). Elsevier Publications, p. 106, 2013.[Abstract]


Peptidoglycan (PG) hydrolases or autolysins are a group of enzymes which degrades bacterial cell wall at specific sites. Staphylococcus aureus produces two major PG hydrolases: major autolysin (Atl) and Aaa, a autolysin/adhesin protein. Amidase are surface-associated proteins that have both enzymatic and adhesive functions. AtlA is the most predominant autolysin in Staphylococcus aureus. The AltA protein only with an amidase domain and the two repeat sequences R1 and R2 is 62 kDa. Another autolysin/ adhesin present on the cell surface of S. aureus (Aaa) is a 35 kDa protein containing two direct LysM (lysine motif) repeats at the N-terminal and catalytic domain in the C-terminus. The amidase domain of the atlA gene with two repeat regions (amiE-R1,2) was cloned and expressed as N-terminal His-tag fusion protein in pQE30 (Qiagen) vector in E.coli M15 (Biswas et al., 2006). For the over-expression of His-tag Aaa; the aaa open reading frame was cloned in the pET28 (Novagen) vector designed to express proteins as fusions with a His6-tag at the N-terminus (Biswas R, 2006). We grew the E. coli M15 cells carrying the altA gene encoding the His-tagged protein in LB medium containing ampicillin to an OD600 nm of 0.5; expression was then induced with different concentrations of isopropyl β-D-thiogalactoside (IPTG). Cells were harvested by centrifugation, and lysed by freeze thaw method with the addition of lysozyme. Proteins were purified under denaturing conditions with 8M urea and analysed by 10 % SDS PAGE. In the same way, we also cultured the E. coli BL21 cells carrying the aaa gene but in LB medium with kanamycin instead of ampicillin. The optimal IPTG concentration for both was found to be 1mM. Isolation of insoluble and soluble fractions of recombinant protein from the bacterial cell lysate and their analysis confirmed the presence of both the recombinant proteins in the insoluble fraction possibly as inclusion bodies. Large scale purification of both the recombinant proteins was done using nickel (Ni-NTA) affinity chromatography. Maximum protein elution was found in 250mM imidazole buffer in both the cases. Refolding was optimized in refolding buffer containing 0.5mM reduced glutathione: 0.5mM oxidised glutathione (1:1) in 50mM Tris HCl pH-8.0. After refolding the bioactivity of both the proteins were assayed. Cell lysis activity of the refolded protein was observed to be more for 61KDa protein than the 35KDa protein. The refolded 61kDa amidase was found to be strongly binding to fibronectin found by affinity column chromatography.

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Publication Type: Journal Article

Year of Publication Title

2019

Divya Nedungadi, Anupama Binoy, Vinod, V., Muralidharan Vanuopadath, Sudarslal Sadasivan Nair, Dr. Bipin G. Nair, and Dr. Nandita Mishra, “Ginger extract activates caspase independent paraptosis in cancer cells via ER stress, mitochondrial dysfunction, AIF translocation and DNA damage”, Nutrition and Cancer, pp. 1-13, 2019.[Abstract]


AbstractThe rhizome of ginger (Zingiber officinale) a common culinary agent is also known for its medicinal activity. We have earlier reported that pure 6-shogaol, an important component of ginger induces paraptosis in triple negative breast cancer (MDA-MB-231) and non small cell lung (A549) cancer cells. However, the chemopreventive potential of the whole ginger extract in food remains to be elucidated. Here, we demonstrate for the first time that ginger extract (GE) triggers similar anticancer activity/paraptosis against the same cell lines but through different molecular mechanisms. Q-TOF LC-MS analysis of the extract showed the presence of several other metabolites along with 6-shogaol and 6-gingerol. GE induces cytoplasmic vacuolation through ER stress and dilation of the ER. Drastic decrease in the mitochondrial membrane potential and ATP production along with the excess generation of ROS contributed to mitochondrial dysfunction. Consequently, GE caused the translocation of apoptosis inducing factor to the nucleus leading to the fragmentation of DNA. Taken together, these show a novel mechanism for ginger extract induced cancer cell death that can be of potential interest for cancer preventive strategies.

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2019

Anupama Binoy, Divya Nedungadi, Katiyar, N., Chinchu Bose, Dr. Sahadev Shankarappa, Dr. Bipin G. Nair, and Dr. Nandita Mishra, “Plumbagin induces paraptosis in cancer cells by disrupting the sulfhydryl homeostasis and proteasomal function”, Chemico-Biological Interactions, p. 108733, 2019.[Abstract]


Plumbagin (PLB) is an active secondary metabolite extracted from the roots of Plumbago rosea. In this study, we report that plumbagin effectively induces paraptosis by triggering extensive cytoplasmic vacuolation followed by cell death in triple negative breast cancer cells (MDA-MB-231), cervical cancer cells (HeLa) and non-small lung cancer cells (A549) but not in normal lung fibroblast cells (WI-38). The vacuoles originated from the dilation of the endoplasmic reticulum (ER) and were found to be empty. The cell death induced by plumbagin was neither apoptotic nor autophagic. Plumbagin induced ER stress mainly by inhibiting the chymotrypsin-like activity of 26S proteasome as also evident from the accumulation of polyubiquitinated proteins. The vacuolation and cell death were found to be independent of reactive oxygen species generation but was effectively inhibited by thiol antioxidant suggesting that plumbagin could modify the sulfur homeostasis in the cellular milieu. Plumbagin also resulted in a decrease in mitochondrial membrane potential eventually decreasing the ATP production. This is the first study to show that Plumbagin induces paraptosis through proteasome inhibition and disruption of sulfhydryl homeostasis and thus further opens up the lead molecule to potential therapeutic strategies for apoptosis-resistant cancers.

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2017

N. Velusamy, Anupama Binoy, Bobba, K. Naidu, Divya Nedungadi, Dr. Nandita Mishra, and Bhuniya, S., “A bioorthogonal fluorescent probe for mitochondrial hydrogen sulfide: new strategy for cancer cell labeling”, Chem Commun (Camb)., vol. 53, no. 62, pp. 8802-8805, 2017.[Abstract]


We report the application of a chemodosimeter {'}turn on{'} fluorescent probe for detecting endogenous H2S formation in cancer cells. Mito-HS showed a bathochromic shift in the UV-vis-absorption spectrum from 355 nm to 395 nm in the presence of H2S. Furthermore{,} it showed an [similar]43-fold fluorescence enhancement at [small lambda]em = 450 nm in the presence of H2S (200 [small mu ]M). The cancer cell-specific fluorescence imaging reveals that Mito-HS has the ability to distinguish cancer cells from normal cells based on the level of endogenous H2S formation. In due course{,} Mito-HS would be a powerful cancer biomarker based on its ability to estimate endogenous H2S formation in living cells.

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2016

Y. K. Yasoda, Bobba, K. Naidu, Divya Nedungadi, Dutta, D., M. Kumar, S., Kothurkar, N., Mishra, N., and Bhuniya, S., “GSH-responsive biotinylated poly(vinyl alcohol)-grafted GO as a nanocarrier for targeted delivery of camptothecin”, RSC Adv., vol. 6, pp. 62385-62389, 2016.[Abstract]


A water-soluble and biocompatible polymer{,} i.e. biotinylated poly(vinyl alcohol)-grafted graphene oxide (GO){,} was used as a nanocarrier for targeted delivery of anticancer drug camptothecin (CPT). The extent of CPT release in the presence of glutathione (GSH) from GO-biotinPVA-CPT was monitored by the increase in the fluorescence intensity{,} at [small lambda]max = 450 nm. The cell-specific (HeLa) antiproliferative activity of GO-biotinPVA-CPT makes it suitable to be used for targeted delivery of chemotherapeutics to cancerous cells. More »»

Publication Type: Patent

Year of Publication Title

2018

S. Bhuniya, Dr. Nandita Mishra, Velusamy, N., Anupama Binoy, Bobba, K. Naidu, and Divya Nedungadi, “Flourescent Exomarker Probes for Hydrogen Sulfide Detection”, U.S. Patent 15 / 956 , 4742018.[Abstract]


A fluorescence probe with mitochondrial targeting and two-photon property, its preparation method and application in detecting and tracking endogenous H2S in samples or living cells. The fluorescent probe is prepared by a four-step preparation method and demonstrates a UV-vis absorption increment λab=395 nm and ˜43 fold higher fluorescence intensity in the presence of H2S. The probe further demonstrates stability, selectivity for H2S over competing agents and sensitivity as low as 20 nm. A method of detecting endogenous H2S rapidly in the absence of any external stimulators is provided. Samples are contacted with the probe and the changes in fluorescence are monitored to detect H2S levels. The disclosed probe is non-toxic and suitable as a biomarker and therapeutic molecule in cancer and other diseases.

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