16 research outputs found
J R SOC INTERFACE
Enzymes are dynamic entities, and their dynamic properties are clearly linked to their biological function. It follows that dynamics ought to play an essential role in enzyme evolution. Indeed, a link between conformational diversity and the emergence of new enzyme functionalities has been recognized for many years. However, it is only recently that state-of-the-art computational and experimental approaches are revealing the crucial molecular details of this link. Specifically, evolutionary trajectories leading to functional optimization for a given host environment or to the emergence of a new function typically involve enriching catalytically competent conformations and/or the freezing out of non-competent conformations of an enzyme. In some cases, these evolutionary changes are achieved through distant mutations that shift the protein ensemble towards productive conformations. Multifunctional intermediates in evolutionary trajectories are probably multi-conformational, i.e. able to switch between different overall conformations, each competent for a given function. Conformational diversity can assist the emergence of a completely new active site through a single mutation by facilitating transition-state binding. We propose that this mechanism may have played a role in the emergence of enzymes at the primordial, progenote stage, where it was plausibly promoted by high environmental temperatures and the possibility of additional phenotypic mutationsDepartment of Chemistry, BMC, Uppsala University, Box 576, 751 23 Uppsala, Sweden
Departamento de Quimica Fisica, Facultad de Ciencias, University of Granada, 18071 Granada, SpainWallenberg Academy Fellowship to S.C.L.K. from the Knut and Alice Wallenberg Foundation (KAW 2013.0124)Grant RGP0041/2017 from the Human Frontier Science ProgramFEDER Funds and grant BIO2015–66426-R to J.M.S.R. from the Spanish Ministry of Economy and Competitivenes
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G-Protein coupled receptors: structure and function in drug discovery
The G-protein coupled receptors (GPCRs) superfamily comprise similar proteins arranged into families or classes thus making it one of the largest in the mammalian genome. GPCRs take part in many vital physiological functions making them targets for numerous novel drugs. GPCRs share some distinctive features, such as the seven transmembrane domains, they also differ in the number of conserved residues in their transmembrane domain. Here we provide an introductory and accessible review detailing the computational advances in GPCR pharmacology and drug discovery. An overview is provided on family A-C GPCRs; their structural differences, GPCR signalling, allosteric binding and cooperativity. The dielectric constant (relative permittivity) of proteins is also discussed in the context of site-specific environmental effects
Cooperativity and flexibility in enzyme evolution
Enzymes are flexible catalysts, and there has been substantial
discussion about the extent to which this flexibility contributes
to their catalytic efficiency. What has been significantly less
discussed is the extent to which this flexibility contributes to
their evolvability. Despite this, recent years have seen an
increasing number of both experimental and computational
studies that demonstrate that cooperativity and flexibility play
significant roles in enzyme innovation. This review covers key
developments in the field that emphasize the importance of
enzyme dynamics not just to the evolution of new enzyme
function(s), but also as a property that can be harnessed in the
design of new artificial enzymes.The European Research Council has provided financial support under the
European Community’s Seventh Framework Program (FP7/2007-2013)/ERC
Grant Agreement No. 306474. This work was also funded by the Feder
Funds, Grants from the Spanish Ministry of Economy and Competitiveness
(BIO2015-66426-R and CSD2009-00088) and the Human Frontier Science
Program (RGP0041/2017). A.P. is a Wenner-Gren Foundations Postdoctoral
Fellow and S. C. L. K. is a Wallenberg Academy Fellow
Mechanisms of Intramolecular Communication in a Hyperthermophilic Acylaminoacyl Peptidase: A Molecular Dynamics Investigation
Protein dynamics and the underlying networks of intramolecular interactions and communicating residues within the three-dimensional (3D) structure are known to influence protein function and stability, as well as to modulate conformational changes and allostery. Acylaminoacyl peptidase (AAP) subfamily of enzymes belongs to a unique class of serine proteases, the prolyl oligopeptidase (POP) family, which has not been thoroughly investigated yet. POPs have a characteristic multidomain three-dimensional architecture with the active site at the interface of the C-terminal catalytic domain and a β-propeller domain, whose N-terminal region acts as a bridge to the hydrolase domain. In the present contribution, protein dynamics signatures of a hyperthermophilic acylaminoacyl peptidase (AAP) of the prolyl oligopeptidase (POP) family, as well as of a deletion variant and alanine mutants (I12A, V13A, V16A, L19A, I20A) are reported. In particular, we aimed at identifying crucial residues for long range communications to the catalytic site or promoting the conformational changes to switch from closed to open ApAAP conformations. Our investigation shows that the N-terminal α1-helix mediates structural intramolecular communication to the catalytic site, concurring to the maintenance of a proper functional architecture of the catalytic triad. Main determinants of the effects induced by α1-helix are a subset of hydrophobic residues (V16, L19 and I20). Moreover, a subset of residues characterized by relevant interaction networks or coupled motions have been identified, which are likely to modulate the conformational properties at the interdomain interface
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Computational investigation of ligand binding of flavonoids in cytochrome P450 receptors
Higher-order epistasis shapes the fitness landscape of a xenobiotic-degrading enzyme
Characterizing the adaptive landscapes that encompass the emergence of novel enzyme functions can provide molecular
insights into both enzymatic and evolutionary mechanisms. Here, we combine ancestral protein reconstruction with biochemical,
structural and mutational analyses to characterize the functional evolution of methyl-parathion hydrolase (MPH), an
organophosphate-degrading enzyme. We identify five mutations that are necessary and sufficient for the evolution of MPH
from an ancestral dihydrocoumarin hydrolase. In-depth analyses of the adaptive landscapes encompassing this evolutionary
transition revealed that the mutations form a complex interaction network, defined in part by higher-order epistasis, that constrained
the adaptive pathways available. By also characterizing the adaptive landscapes in terms of their functional activities
towards three additional organophosphate substrates, we reveal that subtle differences in the polarity of the substrate substituents
drastically alter the network of epistatic interactions. Our work suggests that the mutations function collectively to
enable substrate recognition via subtle structural repositioning.N.T. and E.B.-B. thank the Human Frontier Science
Program (HFSP) for support via research grant RGP0006/2013. N.T. acknowledges
support by the Natural Sciences and Engineering Research Council of Canada (NSERC)
via discovery grants RGPIN 418262-12 and RGPIN 2017-04909. N.T. is a CIHR new
investigator and a Michael Smith Foundation of Health Research (MSFHR) career investigator. S.C.L.K. thanks the Knut and Alice Wallenberg Foundation (Wallenberg
Academy Fellowships 2013.0124 and 2018.0140) and the Swedish National
Infrastructure for Computing (SNIC). D.W.A. thanks NSERC and the MSFHR for
post-doctoral support
Evolution of chalcone isomerase from a noncatalytic ancestor
The emergence of catalysis in a noncatalytic protein scaffold is a rare, unexplored event. Chalcone isomerase (CHI), a key enzyme in plant flavonoid biosynthesis, is presumed to have evolved from a nonenzymatic ancestor related to the widely distributed fatty-acid binding proteins (FAPs) and a plant protein family with no isomerase activity (CHILs). Ancestral inference supported the evolution of CHI from a protein lacking isomerase activity. Further, we identified four alternative founder mutations, i.e., mutations that individually instated activity, including a mutation that is not phylogenetically traceable. Despite strong epistasis in other cases of protein evolution, CHI's laboratory reconstructed mutational trajectory shows weak epistasis. Thus, enantioselective CHI activity could readily emerge despite a catalytically inactive starting point. Accordingly, X-ray crystallography, NMR, and molecular dynamics simulations reveal reshaping of the active site toward a productive substrate-binding mode and repositioning of the catalytic arginine that was inherited from the ancestral fatty-acid binding proteins