A5071298784- Receptor Mechanisms and Signaling
- Protein Kinase Regulation and GTPase Signaling
- Protein Structure and Dynamics
- Enzyme Structure and Function
- Neuroscience and Neuropharmacology Research
- Mass Spectrometry Techniques and Applications
- Ion channel regulation and function
- Supramolecular Self-Assembly in Materials
- Monoclonal and Polyclonal Antibodies Research
- Computational Drug Discovery Methods
- Advanced Fluorescence Microscopy Techniques
- Retinal Development and Disorders
- Cellular transport and secretion
- Viral Infectious Diseases and Gene Expression in Insects
- Photoreceptor and optogenetics research
- Heat shock proteins research
- Cell Image Analysis Techniques
- Erythrocyte Function and Pathophysiology
- Signaling Pathways in Disease
- Metabolism, Diabetes, and Cancer
- Nicotinic Acetylcholine Receptors Study
- Melanoma and MAPK Pathways
- Glycosylation and Glycoproteins Research
- Force Microscopy Techniques and Applications
- Lipid Membrane Structure and Behavior
Stanford University
2020-2024
University of Rochester Medical Center
2014-2022
University of Michigan
2018
University of Rochester
2014-2017
Summary G protein-coupled receptors (GPCRs) activate heterotrimeric proteins by stimulating the exchange of guanine nucleotide in Gα subunit. To visualize this mechanism, we developed a time-resolved cryo-EM approach that examines progression ensembles pre-steady-state intermediates GPCR-G protein complex. Using variability analysis to monitor transitions stimulatory Gs complex with β 2 -adrenergic receptor (β AR) at short sequential time points after GTP addition, identified conformational...
Many chaperones promote nascent polypeptide folding followed by substrate release through ATP-dependent conformational changes. Here we show cryoEM structures of Gα subunit intermediates in complex with full-length Ric-8A, a unique chaperone-client system which is facilitated guanine nucleotide binding to the client G protein. The Ric-8A-Gαi and Ric-8A-Gαq complexes reveal that chaperone employs its extended C-terminal region cradle Ras-like domain Gα, positioning Ras core contact Ric-8A...
Phosphorylation of evolutionarily conserved sites enhances the function Gα subunit chaperone and regulator Ric-8A.
Summary Cryogenic electron microscopy (cryo-EM) has widened the field of structure-based drug discovery by allowing for routine determination membrane protein structures previously intractable. However, despite representing one largest classes therapeutic targets, most inactive-state G protein-coupled receptors (GPCRs) have remained inaccessible cryo-EM because their small size and membrane-embedded nature impedes projection alignment high-resolution map reconstructions. Here we demonstrate...
Inhibitor-2 (I-2) is a prototypic inhibitor of protein phosphatase-1 (PP1), major serine-threonine phosphatase that regulates synaptic plasticity and learning memory. Although I-2 potent PP1 in vitro, our previous work has elucidated that, vivo, may act as positive regulator PP1. Here we show PP1γ, but not PP1α, positively regulate transmission hippocampal neurons. Moreover, demonstrated enhanced PP1γ interaction with its scaffold, neurabin, by Förster resonance energy transfer...
Abstract Inhibitor-2 (I-2) is a prototypic inhibitor of protein phosphatase-1 (PP1), major serine-threonine phosphatase that regulates synaptic plasticity and learning memory. Although I-2 potent PP1 in vitro, our previous work has elucidated that, vivo, may act as positive regulator PP1. Here we show PP1γ, but not PP1α, positively regulate transmission hippocampal neurons. Moreover, demonstrated enhances PP1γ interaction with its scaffold, neurabin, by Förster resonance energy transfer...
Mammalian Ric-8 chaperones regulate the folding and cellular abundance of heterotrimeric G protein α subunits. Ric-8A acts as a chaperone for Gαi/o, Gα12/13 Gαq/11 subunits, while Ric-8B facilitates Gαs/olf folding. No studies have yet delineated precise elements that contribute to this specificity. Here, we determined cryoEM structures complexed Gαs Gαolf, revealing isoform differences in relative positioning contacts between C-terminal α5 helix Gα within concave pocket formed by α-helical...
Abstract GABA (γ-aminobutyric acid) stimulation of the metabotropic B receptor results in prolonged inhibition neurotransmission that is central to brain physiology. belongs Family C G protein-coupled receptors (GPCRs), which operate as dimers relay synaptic neurotransmitter signals into a cellular response through binding and activation heterotrimeric proteins. , however, unique its function an obligate heterodimer agonist protein take place on distinct subunits. Here we show structures...
Ric‐8A is required for the biosynthetic folding of Gαi/q/13. also a guanine nucleotide exchange factor (GEF) that stimulates in vitro Gα subunit GTP binding. Although crucial folding, and therefore, proper G protein heterotrimer function, mechanisms by which itself regulated have yet to be elucidated. We experimentally identified multiple stoichiometrically phosphorylated sites recombinant produced S f 9 cells demonstrate endogenous mammalian protein. Recombinant was enzymatically...
Heterotrimeric G-protein signaling aberrancies contribute to a broad spectrum of diseases including cancers, heart disease, neurodegenerative disorders, and endocrine diseases. Hence the correct folding G protein α subunits is requisite proper coupled receptor signal transduction. Our lab found Ric-8A Ric-8B, be essential genes that encode chaperones for Gα i/q/13 s/olf subunit classes, respectively. In Ric-8 gene deletion or depletion models, do not obtain functional folds, they fail become...
G‐protein coupled receptor (GPCR) signaling is broadly utilized across physiologic systems and the aberrant of pathways contributes to pathologies including cardiovascular, neurodegenerative, endocrine diseases. The correct folding heterotrimeric G protein α subunits requisite for GPCR signal transduction. Our lab identified Ric‐8A Ric‐8B as necessary biosynthetic chaperones i/q/13 Gα s/olf subunit classes, respectively. In Ric‐8 gene deletion or depletion models are folded improperly, fail...