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
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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
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References
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Journal ArticleDOI
Regulation of RAF Activity by 14-3-3 Proteins RAF KINASES ASSOCIATE FUNCTIONALLY WITH BOTH HOMO- AND HETERODIMERIC FORMS OF 14-3-3 PROTEINS
Andreas Fischer,Angela Baljuls,Joerg Reinders,Elena Nekhoroshkova,Claudia Sibilski,Renate Metz,Stefan Albert,Krishnaraj Rajalingam,Mirko Hekman,Ulf R. Rapp +9 more
TL;DR: It is found that B-RAF associates in vivo with 14-3-3 at much higher diversity than A- and C- RAF, and that A-, B-, and C -RAF activity is differentially regulated by its C-terminal and internal 14- 3-3 binding domain.
Journal ArticleDOI
Surface localized and extracellular Glyceraldehyde-3-phosphate dehydrogenase of Bacillus anthracis is a plasminogen binding protein
TL;DR: The presence of a prominent moonlighting enzyme, Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on the surface and in the extracellular medium of Bacillus anthracis and human plasminogen predominantly interacted with the rGapA isoform at physiological concentrations and the interaction was found to be lysine dependent.
Journal ArticleDOI
Activation of protein kinase C by purified bovine brain 14-3-3: comparison with tyrosine hydroxylase activation.
TL;DR: The activation of PKC by 14‐3‐3 is independent of phosphatidylserine and calcium and, as such, is an alternative mechanism for the activation ofPKC that obviates its translocation to membranes.
Journal ArticleDOI
Stabilizer-Guided Inhibition of Protein-Protein Interactions.
Lech-Gustav Milroy,Maria Bartel,Morkos A. Henen,Seppe Leysen,Joris M. C. Adriaans,Luc Brunsveld,Isabelle Landrieu,Christian Ottmann,Christian Ottmann +8 more
TL;DR: This work sets a precedent for the rational design of PPI inhibitors guided by PPI stabilizer-protein complexes while potentially enabling access to new synthetically tractable stabilizers of 14-3-3 and other PPIs.
Journal ArticleDOI
ANIA: ANnotation and Integrated Analysis of the 14-3-3 interactome
TL;DR: The dimeric 14-3-3 proteins dock onto pairs of phosphorylated Ser and Thr residues on hundreds of proteins, and thereby regulate many events in mammalian cells, and to facilitate global analyses of these interactions, a web resource is developed, named ANIA: ANnotation and Integrated Analysis of the 14- 3-3 interactome, which integrates multiple data sets on 14- third-binding phosphoproteins.