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dc.creatorCournia, Zoe
dc.creatorGkeka, Paraskevi
dc.date2017-04-27
dc.date.accessioned2017-05-04T18:06:32Z
dc.date.available2017-05-04T18:06:32Z
dc.identifier.urihttp://hdl.handle.net/21.15102/VISEEM-173
dc.descriptionThis dataset contains Molecular Dynamics simulations trajectories of the normal (wild-type, WT) PI3Ka and the mutant H1047R PI3Ka. We provide two different datasets: 1) MD simulations of only the catalytic subunit of PI3Ka (p110a), and without the regulatory subunit (p85a), in the WT and mutant H1047R forms (PI3Ka-with-p110a-without-p85a. These simulations are given in five independent replicate trajectories. The input and output files are in NAMD format. These simulations are described in Gkeka et al: http://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1003895 2) MD simulations of both the catalytic subunit of PI3Ka (p110a) and the regulatory subunit (p85a), in the WT and mutant H1047R forms (PI3Ka-with-p110a-with-p85a). One simulation set is performed in complex with the PI3Ka inhibitor, PIK-108, and another simulation set without an inhibitor (apo form). The apo forms are provided in both double and single precision for comparison. These simulations are described in Gkeka et al: http://pubs.acs.org/doi/abs/10.1021/jp506423e The input and output files are in GROMACS format.
dc.description.abstractThe PIK3CA gene is one of the most frequently mutated oncogenes in human cancers. It encodes p110a the catalytic subunit of phosphatidylinositol 3-kinase alpha (PI3Ka which activates signaling cascades leading to cell proliferation, survival, and cell growth. The most frequent mutation in PIK3CA is H1047R, which results in enzymatic overactivation. Understanding how the H1047R mutation causes the enhanced activity of the protein in atomic detail is central to developing mutant-specific therapeutics for cancer. To this end, Surface Plasmon Resonance (SPR) experiments and Molecular Dynamics (MD) simulations were carried out for both wild-type (WT) and H1047R mutant p110a proteins. An expanded positive charge distribution on the membrane binding regions of the mutant with respect to the WT protein is observed through MD simulations, which justifies the increased ability of the mutated protein variant to bind to membranes rich in anionic lipids in our SPR experiments. Our results further support an auto-inhibitory role of the C-terminal tail in the WT protein, which is abolished in the mutant protein due to loss of crucial intermolecular interactions. Moreover, Functional Mode Analysis reveals that the H1047R mutation alters the twisting motion of the N-lobe of the kinase domain with respect to the C-lobe and shifts the position of the conserved P-loop residues in the vicinity of the active site. These findings demonstrate the dynamical and structural differences of the two proteins in atomic detail and propose a mechanism of overactivation for the mutant protein. The results may be further utilized for the design of mutant-specific PI3Ka inhibitors that exploit the altered mutant conformation. http://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1003895 Allosteric modulators offer a novel approach for kinase inhibition because they target less conserved binding sites compared to the active site; thus, higher selectivity may be obtained. PIK-108, a known pan phosphoinositide 3-kinase (PI3K) inhibitor, was recently detected to occupy a non-ATP binding site in the PI3Ka C-lobe. This newly identified pocket is located close to residue 1047, which is frequently mutated in human cancers (H1047R). In order to assess the interactions, stability, and any possible allosteric effects of this inhibitor on PI3Ka, extensive molecular dynamics (MD) simulations in aqueous solution were performed for the wild type (WT) human, WT murine, and H1047R human mutant PI3Ka proteins with PIK-108 placed in both catalytic and non-ATP sites. We verify the existence of the second binding site in the vicinity of the hotspot H1047R PI3Ka mutation through binding site identification and MD simulations. PIK-108 remains stable in both sites in all three variants throughout the course of the simulations. We demonstrate that the pose and interactions of PIK-108 in the catalytic site are similar in the murine WT and human mutant forms, while they are significantly different in the case of human WT PI3Ka protein. PIK-108 binding in the non-ATP pocket also differs significantly among the three variants. Finally, we examine whether the non-ATP binding site is implicated in PI3Ka allostery in terms of its communication with the active site using principal component analysis and perform in vitro experiments to verify our hypotheses. http://pubs.acs.org/doi/abs/10.1021/jp506423e
dc.description.sponsorshipVI-SEEM
dc.publisherBRFAA
dc.subjectcancer, PI3Ka, PI3K alpha, Molecular Dynamics simulations, mutation, H1047R, oncogene, breast cancer
dc.titleInvestigating the Structure and Dynamics of the PIK3CA Wild-Type and H1047R Oncogenic Mutant
dc.rights.licenseFree upon citing the publications http://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1003895 http://pubs.acs.org/doi/abs/10.1021/jp506423e


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