Structure of GPCRs and G proteins

Slides:

Advertisements

Similar presentations

Heterotrimeric G proteins and the role of lipids in signaling John Sondek, Ph.D. Depts. of Pharmacology and Biochemistry & Biophyscis.

Advertisements

Rhodopsin Christen Eberhart. Rhodopsin Sequence The Eye Rhodopsin is found in the rods that are located in the eye Rods are composed of stacked disks.

G Proteins - Part 2 Biochemistry 4000 Dr. Ute Kothe.

Crystal Structure of Rhodopsin : A G Protein-Coupled Receptor Authors : Krzysztof Palczewski, Takasi Kumasaka, Tetsuya Hori, Craig A. Behnke, Hiroyuki.

Photoreception: mechanisms of transduction, and non- vertebrate eyes The energy detected:

Structure of Rod and cones cells in retinas Dr. Samina Haq Dept of Biochemistry King Saud University.

Cell Signaling (Lecture 2). Types of signaling Autocrine Signaling Can Coordinate Decisions by Groups of Identical Cells Cells send signals to other.

Monomeric GPCR as a Functional Unit Marc Chabre and Marc le Maire Presented by : Naveena Sivaram.

By Siavoush Dastmalchi Tabriz University of Medical Sciences Tabriz-Iran Modelling the Structures of G Protein-Coupled Receptors Aided by Three-Dimensional.

Caffeine Signaling via Ligand-Receptor Binding Agonist - ligand binding to a receptor and eliciting a response Antagonist - ligand binding to a receptor.

Cell Signaling (Lecture 2)

By: Jillian Rainville Faculty Sponsor: Linda Hufnagel

Cell Signaling (BIO-203) Lecture 4. How the signaling terminates The G α -GTP state is short-lived because the bound GTP is hydrolyzed to GDP in minutes.

STRUCTURAL DYNAMICS OF RHODOPSIN: A G-PROTEIN-COUPLED RECEPTOR by Basak Isin.

Structure of the Rho Family GTP-Binding Protein Cdc42 in Complex with the Multifunctional Regulator RhoGDI Gregory R. Hoffman, Nicolas Nassar, Richard.

3-Dimensional structure of membrane-bound coagulation factor VIII: modeling of the factor VIII heterodimer within a 3-dimensional density map derived by.

Transmembrane and GPCR Mohammed Mohammed Khan PhD Scholar- Department of Biochemistry King Abdul-Aziz University.

Illustrated by: Carrie Wade & Esther Torres

Conformational changes in rhodopsin Example Lecture

Volume 25, Issue 8, Pages e4 (August 2017)

Structure of the Rab7:REP-1 Complex

Sebastian Meyer, Raimund Dutzler Structure

Kristopher Josephson, Naomi J. Logsdon, Mark R. Walter Immunity

Structure of an LDLR-RAP Complex Reveals a General Mode for Ligand Recognition by Lipoprotein Receptors Carl Fisher, Natalia Beglova, Stephen C. Blacklow

Volume 20, Issue 8, Pages (August 2012)

Structural Basis of Rho GTPase-Mediated Activation of the Formin mDia1

Structural Analysis of Engineered Bb Fragment of Complement Factor B

The crystal structure of rhodopsin.

Volume 5, Issue 4, Pages (April 2004)

Yvonne Groemping, Karine Lapouge, Stephen J. Smerdon, Katrin Rittinger

Opioid Receptor Three-Dimensional Structures from Distance Geometry Calculations with Hydrogen Bonding Constraints Irina D. Pogozheva, Andrei L. Lomize,

HyeongJun Kim, Jen Hsin, Yanxin Liu, Paul R. Selvin, Klaus Schulten

Mark A Wall, Bruce A Posner, Stephen R Sprang Structure

Volume 11, Issue 11, Pages (November 2003)

Crystal Structure of ARF1•Sec7 Complexed with Brefeldin A and Its Implications for the Guanine Nucleotide Exchange Mechanism Elena Mossessova, Richard.

Crystal Structures of Ral-GppNHp and Ral-GDP Reveal Two Binding Sites that Are Also Present in Ras and Rap Nathan I. Nicely, Justin Kosak, Vesna de Serrano,

Volume 4, Issue 5, Pages (November 1999)

The Signaling Pathway of Rhodopsin

Crystal Structure of Recombinant Human Interleukin-22

Arnau Cordomí, Gemma Navarro, María S. Aymerich, Rafael Franco

Volume 4, Issue 11, Pages (November 1996)

Crystal Structure of β-Arrestin at 1.9 Å

Volume 91, Issue 5, Pages (November 1997)

Coiled-Coil Domains of SUN Proteins as Intrinsic Dynamic Regulators

Volume 3, Issue 5, Pages (May 1999)

Antonina Roll-Mecak, Chune Cao, Thomas E. Dever, Stephen K. Burley

Volume 6, Issue 6, Pages (December 2000)

Volume 21, Issue 7, Pages (July 2013)

A Putative Mechanism for Downregulation of the Catalytic Activity of the EGF Receptor via Direct Contact between Its Kinase and C-Terminal Domains Meytal.

Volume 15, Issue 11, Pages (November 2007)

by Anna T. Gres, Karen A. Kirby, Vineet N. KewalRamani, John J

Structural Basis of Rab Effector Specificity

Mechanisms Contributing to T Cell Receptor Signaling and Assembly Revealed by the Solution Structure of an Ectodomain Fragment of the CD3ϵγ Heterodimer

Volume 4, Issue 5, Pages (May 1996)

Structure of the Rho Family GTP-Binding Protein Cdc42 in Complex with the Multifunctional Regulator RhoGDI Gregory R. Hoffman, Nicolas Nassar, Richard.

Volume 110, Issue 6, Pages (September 2002)

NSF N-Terminal Domain Crystal Structure

Crystal Structure of the Human Myeloid Cell Activating Receptor TREM-1

Volume 91, Issue 5, Pages (November 1997)

Volume 13, Issue 5, Pages (May 2005)

Structure of an IκBα/NF-κB Complex

Kristopher Josephson, Naomi J. Logsdon, Mark R. Walter Immunity

Structural Basis of Inward Rectification

Volume 16, Issue 6, Pages (December 2004)

Sabine Pokutta, William I. Weis Molecular Cell

Binding Specificity of Retinal Analogs to Photoactivated Visual Pigments Suggest Mechanism for Fine-Tuning GPCR-Ligand Interactions Sundaramoorthy Srinivasan,

Crystal Structure of β-Arrestin at 1.9 Å

Structural Basis for Activation of ARF GTPase

Morgan Huse, Ye-Guang Chen, Joan Massagué, John Kuriyan Cell

Volume 25, Issue 1, Pages (January 2017)

Presentation transcript:

Structure of GPCRs and G proteins Goal of the lecture: Understanding the structural basis of how a GPCR activates a G protein

Heterotrimeric G protein Pathway Clapham Nature. 1996 Jan 25;379(6563):297-299.

Ribbon Diagram of Rhodopsin Structure Palczewski et al, Science. 2000 Aug 4;289(5480):739-745.

Two dimensional Representation of Rhodopsin Palczewski et al, Science. 2000 Aug 4;289(5480):739-745.

The environment of 11-cis retinal chromophore Palczewski et al, Science. 2000 Aug 4;289(5480):739-745.

Salient features of Rhodopsin Structure Organization of the extracellular region serves as the basis seven-helix bundle arrangement 11-cis retinal holds transmembrane regions in the inactive conformation by interacting with key residues that participate in intra-helical interactions

Activation of Rhodopsin Requirement of rigid-body motion of transmembrane helices for light activation of rhodopsin. Farrens et al, Science. 1996 Nov 1;274(5288):768-770.

Design of the Experiment Mutate all Cys to Ser Bring back Cys of interest Construct double Cys mutants Keep Cys at 139 (helix 3) constant Vary 2nd Cys from 247-252 in helix 6 Put spin label on the Cys EPR spectroscopy Farrens et al, Science. 1996 Nov 1;274(5288):768-770.

EPR spectra of inactive (dark state) shown as red trace and activated (meta-rhodopsin II) shown as yellow trace to study interactions between loops 3 and 6 Farrens et al, Science. 1996 Nov 1;274(5288):768-770.

Results from EPR Spectroscopy Dark State: Distance between Cys at 139 and Cys at 248-251 = 12-14 Å After illumination increases in distances 23-25 Å Conclusion: Helices 3 and 6 move apart from each other after activation

Biochemical Verification of EPR predicted movement of helices Cross link with disulfide reagent, cut with V8 protease Run SDS-PAGE If cross linked 1 band without DTT; 2 bands with DTT Farrens et al, Science. 1996 Nov 1;274(5288):768-770.

Crosslinking of helices 3 and 6 blocks the ability of Rhodopsin to activate Transducin Fluorescence assay to measure GTPgS binding to transducin Farrens et al, Science. 1996 Nov 1;274(5288):768-770.

Conclusions Helix 6 moves with respect to Helix 3 Movement is required for activation of transducin Helix 6 movement causes cytoplasmic loop3 to move Cytoplasmic loop3 is involved in coupling to transducin Farrens et al, Science. 1996 Nov 1;274(5288):768-770.

G protein structure Lambright et al, Nature. 1996 Jan 25;379(6563):311-319.

Space filling model of Ga interacts with Gbg Lambright et al, Nature. 1996 Jan 25;379(6563):311-319.

The Gb interface that interacts with Ga contains key residues required for interaction with effectors Lambright et al, Nature. 1996 Jan 25;379(6563):311-319.

G protein residues involved in regulation of effectors Space filling model of Gbg. Gb is white and Gg is pink. The green region is the area of Gb covered by Ga in the heterotrimer The smaller regions marked by colored dashed lines identify residues involved in interactions with various effectors. Each color corresponds to an effector Ford et al, Science. 1998 May 22;280(5367):1271-1274.

In the heterotrimer the switch II region of Ga is contact with Gb Wall et al, Cell. 1995 Dec 15;83(6):1047-1058.

Changes in the conformation of Ga in the GDP vs GTP bound forms and interactions with Gb GTPgS-Ga Red GDP-Ga Blue Wall et al, Cell. 1995 Dec 15;83(6):1047-1058.

The Switch II region of Ga has different conformation in the GDP and GTP bound states GTPgS GDP Wall et al, Cell. 1995 Dec 15;83(6):1047-1058.

The heterotrimeric G protein interacts with the membrane and receptor Lambright et al, Nature. 1996 Jan 25;379(6563):311-319.

A structural cartoon of G protein interaction with receptor Hamm J Biol Chem. 1998 Jan 9;273(2):669-672.

Evolving view of receptors GPCRs exist as dimers Park et al, Biochemistry. 2004 Dec 21;43(50):15643-15656.

Atomic Force Microscopy Picture of mouse rod-outer segment disc membrane Fotiadis et al, Nature. 2003 Jan 9;421(6919):127-128.

Organization of the cytoplasmic surface of rhodopsin dimers are clearly visible Fotiadis et al, Nature. 2003 Jan 9;421(6919):127-128.

Model of Rhodopsin Dimer Here phosphorylated Rhodopsin is shown binding to arrestin (This would be the Desensitized state) Park et al, Biochemistry. 2004 Dec 21;43(50):15643-15656.

Model of rhodopsin dimer binding to one molecule of transducin Park et al, Biochemistry. 2004 Dec 21;43(50):15643-15656.

Receptor Dimer Activation of G proteins Park et al, Biochemistry. 2004 Dec 21;43(50):15643-15656. A movie of this molecule is available from http://stke.sciencemag.org/cgi/content/full/sigtrans;2005/276/tr10/DC1