Identification of two distinct inactive conformations of the beta2-adrenergic receptor reconciles structural and biochemical observations. Export

@article{citeulike:4230367, abstract = {Fully understanding the mechanisms of signaling proteins such as G protein-coupled receptors (GPCRs) will require the characterization of their conformational states and the pathways connecting those states. The recent crystal structures of the beta(2)- and beta(1)-adrenergic receptors in a nominally inactive state constituted a major advance toward this goal, but also raised new questions. Although earlier biochemical observations had suggested that these receptors possessed a set of contacts between helices 3 and 6, known as the ionic lock, which was believed to form a molecular switch for receptor activation, the crystal structures lacked these contacts. The unexpectedly broken ionic lock has raised questions about the true conformation(s) of the inactive state and the role of the ionic lock in receptor activation and signaling. To address these questions, we performed microsecond-timescale molecular dynamics simulations of the beta(2)-adrenergic receptor (beta(2)AR) in multiple wild-type and mutant forms. In wild-type simulations, the ionic lock formed reproducibly, bringing the intracellular ends of helices 3 and 6 together to adopt a conformation similar to that found in inactive rhodopsin. Our results suggest that inactive beta(2)AR exists in equilibrium between conformations with the lock formed and the lock broken, whether or not the cocrystallized ligand is present. These findings, along with the formation of several secondary structural elements in the beta(2)AR loops during our simulations, may provide a more comprehensive picture of the inactive state of the beta-adrenergic receptors, reconciling the crystal structures with biochemical studies.}, author = {Dror, Ron O. and Arlow, Daniel H. and Borhani, David W. and Jensen, Morten {\O}. and Piana, Stefano and Shaw, David E.}, citeulike-article-id = {4230367}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0811065106}, citeulike-linkout-1 = {http://www.pnas.org/content/106/12/4689.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/106/12/4689.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/19258456}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=19258456}, day = {24}, doi = {10.1073/pnas.0811065106}, issn = {1091-6490}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {activation, b2ar, dynamics, flexibility, md, modeling, structure, theory}, month = {March}, number = {12}, pages = {4689--4694}, posted-at = {2009-10-08 11:13:04}, priority = {2}, title = {Identification of two distinct inactive conformations of the beta2-adrenergic receptor reconciles structural and biochemical observations.}, url = {http://dx.doi.org/10.1073/pnas.0811065106}, volume = {106}, year = {2009} }

TY - JOUR ID - citeulike:4230367 L3 - citeulike-article-id:4230367 N2 - Fully understanding the mechanisms of signaling proteins such as G protein-coupled receptors (GPCRs) will require the characterization of their conformational states and the pathways connecting those states. The recent crystal structures of the beta(2)- and beta(1)-adrenergic receptors in a nominally inactive state constituted a major advance toward this goal, but also raised new questions. Although earlier biochemical observations had suggested that these receptors possessed a set of contacts between helices 3 and 6, known as the ionic lock, which was believed to form a molecular switch for receptor activation, the crystal structures lacked these contacts. The unexpectedly broken ionic lock has raised questions about the true conformation(s) of the inactive state and the role of the ionic lock in receptor activation and signaling. To address these questions, we performed microsecond-timescale molecular dynamics simulations of the beta(2)-adrenergic receptor (beta(2)AR) in multiple wild-type and mutant forms. In wild-type simulations, the ionic lock formed reproducibly, bringing the intracellular ends of helices 3 and 6 together to adopt a conformation similar to that found in inactive rhodopsin. Our results suggest that inactive beta(2)AR exists in equilibrium between conformations with the lock formed and the lock broken, whether or not the cocrystallized ligand is present. These findings, along with the formation of several secondary structural elements in the beta(2)AR loops during our simulations, may provide a more comprehensive picture of the inactive state of the beta-adrenergic receptors, reconciling the crystal structures with biochemical studies. IS - 12 JF - Proceedings of the National Academy of Sciences of the United States of America SN - 1091-6490 EP - 4694 TI - Identification of two distinct inactive conformations of the beta2-adrenergic receptor reconciles structural and biochemical observations. VL - 106 SP - 4689 KW - activation KW - b2ar KW - dynamics KW - flexibility KW - md KW - modeling KW - structure KW - theory AU - Dror, Ron AU - Arlow, Daniel AU - Borhani, David AU - Jensen, Morten AU - Piana, Stefano AU - Shaw, David PY - 2009/03/24/ UR - http://dx.doi.org/10.1073/pnas.0811065106 ER -