Characterization of sulfated polysaccharide activity against virulent Plasmodium falciparum PHISTb/RLP1 protein

Jennifer M Mutisya; Victor A Mobegi; Johnson K Kinyua; Martha N Kivecu; Raphael O Okoth; Gladys C Chemwor; Edwin W Mwakio; Agnes C Cheruiyot; Redempta A Yeda; Charles O Okello; Jackline A Juma; Benjamin H Opot; Dennis W Juma; Amanda L Roth; Hosea M Akala; Ben M Andagalu

doi:10.12688/f1000research.26756.2

Characterization of sulfated polysaccharide activity against virulent Plasmodium falciparum PHISTb/RLP1 protein

F1000Res. 2020 Oct 23:9:1268. doi: 10.12688/f1000research.26756.2. eCollection 2020.

Authors

Jennifer M Mutisya^{1

2}, Victor A Mobegi³, Johnson K Kinyua², Martha N Kivecu¹, Raphael O Okoth¹, Gladys C Chemwor¹, Edwin W Mwakio¹, Agnes C Cheruiyot¹, Redempta A Yeda¹, Charles O Okello¹, Jackline A Juma¹, Benjamin H Opot¹, Dennis W Juma¹, Amanda L Roth¹, Hosea M Akala¹, Ben M Andagalu¹

Affiliations

¹ Department of Emerging and Infectious Diseases (DEID), United States Army Medical Research Directorate-Africa (USAMRD-A), Kenya Medical Research Institute (KEMRI)/Walter Reed Project, Kisumu, Kenya.
² Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya.
³ Department of Biochemistry, University of Nairobi, Nairobi, Kenya.

Abstract

Background: The emergence of artemisinin resistance in South East Asia calls for urgent discovery of new drug compounds that have antiplasmodial activity. Unlike the classical compound screening drug discovery methods, the rational approach involving targeted drug discovery is less cumbersome and therefore key for innovation of new antiplasmodial compounds. Plasmodium falciparum (Pf) utilizes the process of host erythrocyte remodeling using Plasmodium-helical interspersed sub-telomeric domain (PHIST) containing proteins, which are amenable drug targets. The aim of this study is to identify inhibitors of PHIST from sulfated polysaccharides as new antimalarials. Methods: 251 samples from an ongoing study of epidemiology of malaria and drug resistance sensitivity patterns in Kenya were sequenced for PHISTb/RLP1 gene using Sanger sequencing. The sequenced reads were mapped to the reference Pf3D7 protein sequence of PHISTb/RLP1 using CLC Main Workbench. Homology modeling of both reference and mutant protein structures was achieved using the LOMETs tool. The models were refined using ModRefiner for energy minimization. Ramachandran plot was generated by ProCheck to assess the conformation of amino acids in the protein model. Protein binding sites predictions were assessed using FT SITE software. We searched for prospective antimalarials from PubChem. Docking experiments were achieved using AutoDock Vina and analysis results visualized in PyMOL. Results: Sanger sequencing generated 86 complete sequences. Upon mapping of the sequences to the reference, 12 non-synonymous single nucleotide polymorphisms were considered for mutant protein structure analysis. Eleven drug compounds with antiplasmodial activity were identified. Both modeled PHISTb/RLP1 reference and mutant structures had a Ramachandran score of >90% of the amino acids in the favored region. Ten of the drug compounds interacted with amino acid residues in PHISTb and RESA domains, showing potential activity against these proteins. Conclusion: This research identifies inhibitors of exported proteins that can be used in in vitro tests against the Plasmodium parasite.

Keywords: Anti-malarials; Exported proteins; Interactions; P. falciparum; PHISTb/RLP1; Sulfated polysaccharides.

Grants and funding

Funding for this study was provided by the Armed Forces Health Surveillance Branch (AFHSB) - Global Emerging Infections Surveillance and Response System. This work was also supported through the DELTAS Africa Initiative [DELGEME grant 107740/Z/15/Z]. The DELTAS Africa Initiative is an independent funding scheme of the African Academy of Sciences (AAS)’s Alliance for Accelerating Excellence in Science in Africa (AESA) and supported by the New Partnership for Africa’s Development Planning and Coordinating Agency (NEPAD Agency) with funding from the Wellcome Trust [DELGEME grant 107740/Z/15/Z] and the UK government. The views expressed in this publication are those of the author(s) and not necessarily those of AAS, NEPAD Agency, Wellcome Trust or the UK government. Material has been reviewed by the Walter Reed Army Institute of Research. There is no objection to its presentation and/or publication. The opinions or assertions contained herein are the private views of the author, and are not to be construed as official, or as reflecting true views of the Department of the Army or the Department of Defense. The investigators have adhered to the policies for protection of human subjects as prescribed in AR 70–25.