INFECTIOUS DISEASES / Parasitic Diseases
Understanding malaria parasites biology with an aim to identify new drug and vaccine candidate antigens.
Description of Research
Food Vacuole- Target for new drug discovery against malaria parasite
Our Group has been working on illustration of P. falciparum food vacuole proteome to understand process of hemoglobin degradation as well as to know the proteins that are involved in resistance to current antimalarials. In 2013, we identified a ~200kDa protein complex consisting of falcipain2/2’, plasmepsin II, plasmepsin IV, histo aspartic protease and Heme Detoxification protein (HDP) inside the food vacuole that is involved in hemoglobin (Hb) degradation and hemozoin (Hz) formation. Using recombinant PfHDP and falcipain-2, we developed an in vitro Hb degradation and Hz formation assay. Further using in silico and biochemical approaches, we characterized PfHDP for heme binding as well as hemoglobin binding sites. Screening of compounds from Maybridge Screening Collection that bound the modeled PfHDP structure, identified a compound that inhibited parasite growth in a dose dependent manner, thus paving the way for testing its potential as a new drug candidate. Presently, we are working on this scaffold by computational, structural and experimental approaches for rational drug design to combat drug resistant malaria.
Using proteome approaches, we have recently identified a number of transporters that show frame-shift or missense mutations in resistance field isolates. In collaboration with Dr Asif Mohmmed and Dr Inderjeet Kaur, we are presently developing CRISPR-CAS9 tools to understand the functional relevance of these mutations.
Novel host-parasite interactions involved in Plasmodium falciparum invasion of human RBCs
More than 50 surface antigens are expressed on Plasmodium merozoite surface, however till date ten possible interactions between these antigens and their respective receptors on human RBCs have been characterized. Our group along with other colleagues has recently described two novel erythrocyte receptors; Intercellular Adhesion Molecule 4 (ICAM4) and Cyclophilin B that seem to play important role(s) at different steps of merozoite invasion of human RBCs. Presently work is on to identify novel inhibitor(s), which can block these interactions and possibly the invasion.
Proteome analysis to identify novel protein complexes on Plasmodium merozoite surface and to identify pathways/proteins that are regulated by post-translation modifications
We have been applying proteome-based approaches to illustrate protein complexes on Plasmodium merozoite surface and also to identify pathways/proteins that undergo post-translational modifications. In this context, we have recently described a novel, 6-Cys protein complex comprising of Pfs41, Pfs38, Pfs12, GLURP, SERA5 & MSP-1 and a MSP 3 complex comprising of MSP-1, MSP-6 and MSP-7 proteins. Using liquid chromatography and tandem mass spectrometric analysis, we have first time described lysine as well as arginine methylated Plasmodium proteins at asexual blood stages. Work is on to identify the pathways that are regulated by these proteins and also to develop novel inhibitors to parasite enzymes that are responsible for methylation of parasite proteins.
Deshmukh, A., Chourasia, B.K., Mehrotra, S., Kana, I.H., Paul, G., Panda, A., Kaur, I., Singh, S.K., Rathore, S., Das, A., Gupta, P., Md, K., Ghakar, S.K., Mohmmed, A., Theisen, M., Malhotra, P. 2018. Plasmodium falciparum MSP3 exists in a complex on the merozoite surface and generates antibody response during natural infection. Infect Immun pii: IAI.00067-18. doi: 10.1128/IAI.00067-18 PubMed link
Iyer, G.R., Singh, S., Kaur, I., Agarwal, S., Siddiqui, M.A., Bansal, A., Kumar, G., Saini, E., Paul, G., Mohmmed, A., Chitnis, C.E., Malhotra, P. 2018. Calcium-dependent phosphorylation of <i>Plasmodium falciparum</i> serine repeat antigen 5 triggers merozoite egress. J Biol Chem. 293, 9736-9746. doi: 10.1074/jbc.RA117.001540 PubMed link
Paul, G., Deshmukh, A., Kumar Chourasia, B., Kalamuddin, M., Panda, A., Kumar Singh, S., Gupta, P.K., Mohmmed, A., Chauhan, V.S., Theisen, M., Malhotra, P. 2018. Protein-protein interaction studies reveal the Plasmodium falciparum merozoite surface protein-1 region involved in a complex formation that binds to human erythrocytes. Biochem J 475, 1197-1209. doi: 10.1042/BCJ20180017 PubMed link
Prakash, P., Zeeshan, M., Saini, E., Muneer, A., Khurana, S., Kumar Chourasia, B., Deshmukh, A., Kaur, I., Dabral, S., Singh, N., Anam, Z., Chaurasiya, A., Kaushik, S., Dahiya, P., Kalamuddin, M., Kumar Thakur, J., Mohmmed, A., Ranganathan, A., Malhotra, P. 2017. Human Cyclophilin B forms part of a multi-protein complex during erythrocyte invasion by Plasmodium falciparum. Nat Commun 8, 1548 doi: 10.1038/s41467-017-01638-6 PubMed link
Prakash, P., Zeeshan, M., Saini, E., Muneer, A., Khurana, S., Kumar Chourasia, B., Deshmukh, A., Kaur, I., Dabral, S., Singh, N., Anam, Z., Chaurasiya, A., Kaushik, S., Dahiya, P., Kalamuddin, M., Kumar Thakur, J., Mohmmed, A., Ranganathan, A., Malhotra, P. 2017. Human Cyclophilin B forms part of a multi-protein complex during erythrocyte invasion by Plasmodium falciparum. Nat Commun 8, 1548 doi: 10.1038/s41467-017-01638-6 PubMed linkGupta, P., Mehrotra, S., Sharma, A., Chugh, M., Pandey, R., Kaushik, A., Khurana, S., Srivastava, N., Srivastava, T., Deshmukh, A., Panda, A., Aggarwal, P., Bhavesh, N.S., Bhatnagar, R.K., Mohmmed, A., Gupta, D., Malhotra, P. 2017. Exploring Heme and Hemoglobin Binding Regions of Plasmodium Heme Detoxification Protein for New Antimalarial Discovery. J Med Chem 60, 8298-8308 doi: 10.1021/acs.jmedchem.7b00089 PubMed link
Zeeshan, M., Kaur, I., Joy, J., Saini, E., Paul, G., Kaushik, A., Dabral, S., Mohmmed, A., Gupta, D., Malhotra, P. 2017. Proteomic Identification and Analysis of Arginine-Methylated Proteins of Plasmodium falciparum at Asexual Blood Stages. J Proteome Res. 16, 368-383. doi: 10.1021/acs.jproteome.5b01052 Pubmed link