Allosteric nanobodies reveal the dynamic range and diverse mechanisms of G-protein-coupled receptor activation.
Nature 535(7612):448 (2016)
G-protein-coupled receptors (GPCRs) modulate many physiological processes by transducing a variety of extracellular cues into intracellular responses. Ligand binding to an extracellular orthosteric pocket propagates conformational change to the receptor cytosolic region to promote binding and ac...
Allosteric coupling from G protein to the agonist-binding pocket in GPCRs.
Nature 535(7610):182 (2016)
G-protein-coupled receptors (GPCRs) remain the primary conduit by which cells detect environmental stimuli and communicate with each other. Upon activation by extracellular agonists, these seven-transmembrane-domain-containing receptors interact with heterotrimeric G proteins to regulate downstr...
Crystal structure of the human σ1 receptor.
Nature 532(7600):527 (2016)
The human σ1 receptor is an enigmatic endoplasmic-reticulum-resident transmembrane protein implicated in a variety of disorders including depression, drug addiction, and neuropathic pain. Recently, an additional connection to amyotrophic lateral sclerosis has emerged from studies of human geneti...
High-density grids for efficient data collection from multiple crystals.
Acta Crystallogr D Struct Biol 72(Pt 1):2 (2016)
Higher throughput methods to mount and collect data from multiple small and radiation-sensitive crystals are important to support challenging structural investigations using microfocus synchrotron beamlines. Furthermore, efficient sample-delivery methods are essential to carry out productive fem...
Engineering high-affinity PD-1 variants for optimized immunotherapy and immuno-PET imaging.
PNAS 112(47):E6506 (2015)
Signaling through the immune checkpoint programmed cell death protein-1 (PD-1) enables tumor progression by dampening antitumor immune responses. Therapeutic blockade of the signaling axis between PD-1 and its ligand programmed cell death ligand-1 (PD-L1) with monoclonal antibodies has shown rem...
Structural Insights into the Dynamic Process of β2-Adrenergic Receptor Signaling
Cell 162(6):1431 (2015)
Propagation of conformational changes during μ-opioid receptor activation.
Nature 524(7565):375 (2015)
µ-Opioid receptors (µORs) are G-protein-coupled receptors that are activated by a structurally diverse spectrum of natural and synthetic agonists including endogenous endorphin peptides, morphine and methadone. The recent structures of the μOR in inactive and agonist-induced active states (Huang...
Structural insights into µ-opioid receptor activation.
Nature 524(7565):315 (2015)
Activation of the μ-opioid receptor (μOR) is responsible for the efficacy of the most effective analgesics. To shed light on the structural basis for μOR activation, here we report a 2.1 Å X-ray crystal structure of the murine μOR bound to the morphinan agonist BU72 and a G protein mimetic camel...
SIGNAL TRANSDUCTION. Structural basis for nucleotide exchange in heterotrimeric G proteins.
Science 348(6241):1361 (2015)
G protein-coupled receptors (GPCRs) relay diverse extracellular signals into cells by catalyzing nucleotide release from heterotrimeric G proteins, but the mechanism underlying this quintessential molecular signaling event has remained unclear. Here we use atomic-level simulations to elucidate t...
Structural Insights into the Dynamic Process of β2-Adrenergic Receptor Signaling.
Cell 161(5):1101 (2015)
G-protein-coupled receptors (GPCRs) transduce signals from the extracellular environment to intracellular proteins. To gain structural insight into the regulation of receptor cytoplasmic conformations by extracellular ligands during signaling, we examine the structural dynamics of the cytoplasmi...
A comparison of chemical shift sensitivity of trifluoromethyl tags: optimizing resolution in ¹⁹F NMR studies of proteins.
Journal of Biomolecular NMR 62(1):97 (2015)
The elucidation of distinct protein conformers or states by fluorine ((19)F) NMR requires fluorinated moieties whose chemical shifts are most sensitive to subtle changes in the local dielectric and magnetic shielding environment. In this study we evaluate the effective chemical shift dispersion ...
G Protein-coupled Receptor Kinases of the GRK4 Protein Subfamily Phosphorylate Inactive G Protein-coupled Receptors (GPCRs).
Journal of Biological Chemistry 290(17):10775 (2015)
G protein-coupled receptor (GPCR) kinases (GRKs) play a key role in homologous desensitization of GPCRs. It is widely assumed that most GRKs selectively phosphorylate only active GPCRs. Here, we show that although this seems to be the case for the GRK2/3 subfamily, GRK5/6 effectively phosphoryla...
Goniometer-based femtosecond crystallography with X-ray free electron lasers.
PNAS 111(48):17122 (2014)
The emerging method of femtosecond crystallography (FX) may extend the diffraction resolution accessible from small radiation-sensitive crystals and provides a means to determine catalytically accurate structures of acutely radiation-sensitive metalloenzymes. Automated goniometer-based instrumen...
Covalent agonists for studying G protein-coupled receptor activation.
PNAS 111(29):10744 (2014)
Structural studies on G protein-coupled receptors (GPCRs) provide important insights into the architecture and function of these important drug targets. However, the crystallization of GPCRs in active states is particularly challenging, requiring the formation of stable and conformationally homo...
The role of protein dynamics in GPCR function: insights from the β2AR and rhodopsin.
Current Opinion in Cell Biology 27:136 (2014)
G protein-coupled receptors (GPCRs) are versatile signaling proteins that mediate complex cellular responses to hormones and neurotransmitters. Recent advances in GPCR crystallography have provided inactive and active state structures for rhodopsin and the β2 adrenergic receptor (β2AR). Although...
Activation and allosteric modulation of a muscarinic acetylcholine receptor.
Nature 504(7478):101 (2013)
Despite recent advances in crystallography and the availability of G-protein-coupled receptor (GPCR) structures, little is known about the mechanism of their activation process, as only the β2 adrenergic receptor (β2AR) and rhodopsin have been crystallized in fully active conformations. Here we ...
Applications of molecular replacement to G protein-coupled receptors.
Acta Crystallographica Section D 69(Pt 11):2287 (2013)
G protein-coupled receptors (GPCRs) are a large class of integral membrane proteins involved in regulating virtually every aspect of human physiology. Despite their profound importance in human health and disease, structural information regarding GPCRs has been extremely limited until recently. ...
Adrenaline-activated structure of β2-adrenoceptor stabilized by an engineered nanobody.
Nature 502(7472):575 (2013)
G-protein-coupled receptors (GPCRs) are integral membrane proteins that have an essential role in human physiology, yet the molecular processes through which they bind to their endogenous agonists and activate effector proteins remain poorly understood. So far, it has not been possible to captur...
The role of ligands on the equilibria between functional states of a G protein-coupled receptor.
Journal of the American Chemical Society 135(25):9465 (2013)
G protein-coupled receptors exhibit a wide variety of signaling behaviors in response to different ligands. When a small label was incorporated on the cytosolic interface of transmembrane helix 6 (Cys-265), (19)F NMR spectra of the β2 adrenergic receptor (β2AR) reconstituted in maltose/neopentyl...
Structure of active β-arrestin-1 bound to a G-protein-coupled receptor phosphopeptide.
Nature 497(7447):137 (2013)
The functions of G-protein-coupled receptors (GPCRs) are primarily mediated and modulated by three families of proteins: the heterotrimeric G proteins, the G-protein-coupled receptor kinases (GRKs) and the arrestins. G proteins mediate activation of second-messenger-generating enzymes and other ...