Molecular dynamics simulations and biochemical characterization of Pf14-3-3 and PfCDPK1 interaction towards its role in growth of human malaria parasite
Ravi Jain,Pinki Dey,Sakshi Gupta,Soumya Pati,Arnab Bhattacherjee,Manoj Munde,Shailja Singh,Shailja Singh +7 more
TLDR
Overall, this study characterizes 14-3-3I as a scaffold protein in the malaria parasite and unveils CDPK1 as its previously unidentified target, setting a precedent for the rational design of 14- 3-3 based PPI inhibitors by utilizing 14-2-3 recognition motif peptides, as a potential antimalarial strategy.Abstract:
Scaffold proteins play pivotal role as modulators of cellular processes by operating as multipurpose conformation clamps. 14-3-3 proteins are gold-standard scaffold modules that recognize phosphoSer/Thr (pS/pT) containing conserved motifs, and confer conformational changes leading to modulation of functional parameters of their target proteins. Modulation in functional activity of kinases has been attributed to their interaction with 14-3-3 proteins. Herein, we have annotated and characterized PF3D7_0818200 as 14-3-3 isoform I in Plasmodium falciparum 3D7, and its interaction with one of the key kinases of the parasite, Calcium-Dependent Protein Kinase 1 (CDPK1) by performing various analytical biochemistry and biophysical assays. Molecular dynamics simulation studies indicated that CDPK1 polypeptide sequence (61KLGpS64) behaves as canonical Mode I-type (RXXpS/pT) consensus 14-3-3 binding motif, mediating the interaction. The 14-3-3I/CDPK1 interaction was validated in vitro with ELISA and SPR, which confirmed that the interaction is phosphorylation dependent, with binding affinity constant of 670 ± 3.6 nM. The interaction of 14-3-3I with CDPK1 was validated with well characterized optimal 14-3-3 recognition motifs: Mode I-type ARSHpSYPA and Mode II-type RLYHpSLPA, by simulation studies and ITC. This interaction was found to marginally enhance CDPK1 functional activity. Furthermore, interaction antagonizing peptidomimetics showed growth inhibitory impact on the parasite indicating crucial physiological role of 14-3-3/CDPK1 interaction. Overall, this study characterizes 14-3-3I as a scaffold protein in the malaria parasite and unveils CDPK1 as its previously unidentified target. This sets a precedent for the rational design of 14-3-3 based PPI inhibitors by utilizing 14-3-3 recognition motif peptides, as a potential antimalarial strategy.read more
Citations
More filters
Journal ArticleDOI
Parasite and Host Erythrocyte Kinomics of Plasmodium Infection
TL;DR: In this paper, an overview of the updated kinome of Plasmodium falciparum, the species that is the largest contributor to malaria mortality, and current knowledge pertaining to the function of parasite-encoded protein kinases during the parasite's life cycle is presented.
Journal ArticleDOI
The search for molecular mimicry in proteins carried by extracellular vesicles secreted by cells infected with Plasmodium falciparum.
TL;DR: In this paper, a set of eight proteins with a remarkable resemblance to human proteins were found to be moonlighting proteins carried by Plasmodium falciparum molecules carried by extracellular vesicles.
Journal ArticleDOI
Designing and development of phthalimides as potent anti-tubulin hybrid molecules against malaria.
Vigyasa Singh,Rahul S. Hada,Ravi Jain,Manu Vashistha,Geeta Kumari,Snigdha Singh,Neha Sharma,Meenakshi Bansal,Poonam,Martin Zoltner,Conor R. Caffrey,Brijesh Rathi,Shailja Singh +12 more
TL;DR: In this paper , phthalimide analogs possessing the bioactive scaffolds, benzimidazole and 1,2,3-triazole, were evaluated for in vitro and in vivo anti-plasmodial activity without any apparent hemolysis, or cytotoxicity.
Journal ArticleDOI
Target-Based Virtual Screening of Natural Compounds Identifies a Potent Antimalarial With Selective Falcipain-2 Inhibitory Activity
Amad Uddin,Sonal Gupta,Taj Mohammad,Diksha Shahi,Afzal Hussain,Mohamed F. Alajmi,Hesham R. El-Seedi,Imtaiyaz Hassan,Shailja Singh,Mohammad Abid +9 more
TL;DR: ST72 with CQ resulted in improved growth inhibitory activity than individual drugs in both in vitro and in vivo studies and did not show any significant hemolysis or cytotoxicity against human HepG2 cells suggesting a good safety profile.
Journal ArticleDOI
cAMP-Dependent Signaling Pathways as Potential Targets for Inhibition of Plasmodium falciparum Blood Stages.
Edwin Lasonder,Kunal R. More,Shailja Singh,Malak Haidar,Daniela Bertinetti,Eileen J. Kennedy,Friedrich W. Herberg,Anthony A. Holder,Gordon Langsley,Gordon Langsley,Chetan E. Chitnis +10 more
TL;DR: In this article, the role of signaling pathways in regulation of the key processes of merozoite egress and red blood cell invasion by Plasmodium falciparum and, in particular, the importance of the second messengers, cAMP and Ca2+, and cyclic nucleotide dependent kinases.
References
More filters
Journal ArticleDOI
Regulation of the Regulators: Post-Translational Modifications, Subcellular, and Spatiotemporal Distribution of Plant 14-3-3 Proteins.
TL;DR: This review summarizes current knowledge of 14-3-3 proteins in plants, including 14- 3-3 interactions, regulatory functions, isoform specificity, and post-translational modifications.
Journal ArticleDOI
Genome-wide identification, classification, and expression analysis of 14-3-3 gene family in Populus
TL;DR: The gene structures and their coding protein structures of Populus 14-3-3s are highly conserved among group members, suggesting that members of the same group might also have conserved functions.
Journal ArticleDOI
Differential activation of PKC isozymes by 14-3-3 zeta protein.
TL;DR: The differential activation of various protein kinase C isoforms by 14-3-3 ζ protein is reported, which shows strong positive cooperativity and the classical isozymes show approximately a twofold activation, whereas protein Kinase C ϵ, another novel isozyme, is highly activated.
Journal ArticleDOI
Arabidopsis 14-3-3 epsilon members contribute to polarity of PIN auxin carrier and auxin transport-related development
Jutta Keicher,Nina Jaspert,Katrin Weckermann,Claudia Möller,Christian Throm,Aaron Kintzi,Claudia Oecking +6 more
TL;DR: A fundamental role for the ancient 14-3-3 epsilon group members in regulating PIN polarity and plant development is demonstrated.
Journal ArticleDOI
Structural insights of the MLF1/14-3-3 interaction.
TL;DR: The high‐resolution crystal structure of this binding motif [MLF1(29–42)pSer34] in complex with 14‐3‐3ε is presented and the interaction with isothermal titration calorimetry is analysed.