Dataset Information

Differences in allosteric communication pipelines in the inactive and active states of a GPCR.

ABSTRACT: G-protein-coupled receptors (GPCRs) are membrane proteins that allosterically transduce the signal of ligand binding in the extracellular (EC) domain to couple to proteins in the intracellular (IC) domain. However, the complete pathway of allosteric communication from the EC to the IC domain, including the role of individual amino acids in the pathway is not known. Using the correlation in torsion angle movements calculated from microseconds-long molecular-dynamics simulations, we elucidated the allosteric pathways in three different conformational states of ?2-adrenergic receptor (?2AR): 1), the inverse-agonist-bound inactive state; 2), the agonist-bound intermediate state; and (3), the agonist- and G-protein-bound fully active state. The inactive state is less dynamic compared with the intermediate and active states, showing dense clusters of allosteric pathways (allosteric pipelines) connecting the EC with the IC domain. The allosteric pipelines from the EC domain to the IC domain are weakened in the intermediate state, thus decoupling the EC domain from the IC domain and making the receptor more dynamic compared with the other states. Also, the orthosteric ligand-binding site becomes the initiator region for allosteric communication in the intermediate state. This finding agrees with the paradigm that the nature of the agonist governs the specific signaling state of the receptor. These results provide an understanding of the mechanism of allosteric communication in class A GPCRs. In addition, our analysis shows that mutations that affect the ligand efficacy, but not the binding affinity, are located in the allosteric pipelines. This clarifies the role of such mutations, which has hitherto been unexplained.

SUBMITTER: Bhattacharya S

PROVIDER: S-EPMC4104064 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature

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Differences in allosteric communication pipelines in the inactive and active states of a GPCR.

Bhattacharya Supriyo S Vaidehi Nagarajan N

Biophysical journal 20140701 2

G-protein-coupled receptors (GPCRs) are membrane proteins that allosterically transduce the signal of ligand binding in the extracellular (EC) domain to couple to proteins in the intracellular (IC) domain. However, the complete pathway of allosteric communication from the EC to the IC domain, including the role of individual amino acids in the pathway is not known. Using the correlation in torsion angle movements calculated from microseconds-long molecular-dynamics simulations, we elucidated the ...[more]

PMID: 25028884

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Project description:Metabotropic glutamate receptors (mGluRs) are G protein coupled receptors that play important roles in synaptic plasticity and other neuro-physiological and pathological processes. Allosteric mGluR ligands are particularly promising drug targets because of their modulatory effects--enhancing or suppressing the response of mGluRs to glutamate. The mechanism by which this modulation occurs is not known. Here, we propose the hypothesis that positive and negative modulators will differentially stabilize the active and inactive conformations of the receptors, respectively. To test this hypothesis, we have generated computational models of the transmembrane regions of different mGluR subtypes in two different conformations. The inactive conformation was modeled using the crystal structure of the inactive, dark state of rhodopsin as template and the active conformation was created based on a recent model of the light-activated state of rhodopsin. Ligands for which the nature of their allosteric effects on mGluRs is experimentally known were docked to the modeled mGluR structures using ArgusLab and Autodock softwares. We find that the allosteric ligand binding pockets of mGluRs are overlapping with the retinal binding pocket of rhodopsin, and that ligands have strong preferences for the active and inactive states depending on their modulatory nature. In 8 out of 14 cases (57%), the negative modulators bound the inactive conformations with significant preference using both docking programs, and 6 out of 9 cases (67%), the positive modulators bound the active conformations. Considering results by the individual programs only, even higher correlations were observed: 12/14 (86%) and 8/9 (89%) for ArgusLab and 10/14 (71%) and 7/9 (78%) for AutoDock. These findings strongly support the hypothesis that mGluR allosteric modulation occurs via stabilization of different conformations analogous to those identified in rhodopsin where they are induced by photochemical isomerization of the retinal ligand--despite the extensive differences in sequences between mGluRs and rhodopsin.

Project description:The adenosine A2A receptor (A2AR) belongs to the superfamily of membrane proteins called the G-protein-coupled receptors (GPCRs) that form one of the largest superfamilies of drug targets. Deriving thermostable mutants has been one of the strategies used for crystallization of A2AR in both the agonist and antagonist bound conformational states. The crystal structures do not reveal differences in the activation mechanism of the mutant receptors compared to the wild type receptor, that have been observed experimentally. These differences stem from the dynamic behavior of the mutant receptors. Furthermore, it is not understood how the mutations confer thermostability. Since these details are difficult to obtain from experiments, we have used atomic level simulations to elucidate the dynamic behavior of the agonist and antagonist bound mutants as well the wild type A2AR. We found that significant enthalpic contribution leads to stabilization of both the inactive state (StaR2) and active-like state (GL31) thermostable mutants of A2AR. Stabilization resulting from mutations of bulky residues to alanine is due to the formation of interhelical hydrogen bonds and van der Waals packing that improves the transmembrane domain packing. The thermostable mutant GL31 shows less movement of the transmembrane helix TM6 with respect to TM3 than the wild type receptor. While restricted dynamics of GL31 is advantageous in its purification and crystallization, it could also be the reason why these mutants are not efficient in activating the G proteins. We observed that the calculated stress on each residue is higher in the wild type receptor compared to the thermostable mutants, and this stress is required for activation to occur. Thus, reduced dynamic behavior of the thermostable mutants leading to lowered activation of these receptors originates from reduced stress on each residue. Finally, accurate calculation of the change in free energy for single mutations shows good correlation with the change in the measured thermostability. These results provide insights into the effect of mutations that can be incorporated in deriving thermostable mutants for other GPCRs.

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Differences in allosteric communication pipelines in the inactive and active states of a GPCR.

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Differences in allosteric communication pipelines in the inactive and active states of a GPCR.

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