Rhodopsin: Methods and Protocols

Part I: Historical Overview

1. The G Protein-Coupled Receptor Rhodopsin: A Historical Perspective
Lukas Hofmann and Krzysztof Palczewski

Part II: Rhodopsin Expression, Regeneration and Purification for Structural Studies

2. Rhodopsin Purification from Dark Adapted Bovine Retina
Elise Blankenship and David T. Lodowski

3. Mammalian Expression, Purification, and Crystallization of Rhodopsin Variants
Daniel Mattle, Ankita Singhal, Georg Schmid, Roger Dawson, and Jörg Standfuss

4. Imaging of Rhodopsin Crystals with Two-Photon Microscopy
Grazyna Palczewska and David Salom

Part III: Structure-function characterization of rhodopsin

5. Functional Stability of Rhodopsin in a Bicelle System: Evaluating G-Protein Activation by Rhodopsin in Bicelles
Ali I. Kaya, T. M. Iverson, and Heidi E. Hamm

6. The Rhodopsin-Arrestin-1 Interaction in Bicelles
Qiuyan Chen, Sergey A. Vishnivetskiy, Tiandi Zhuang, Min-Kyu Cho, Tarjani M. Thaker, Charles R. Sanders, Vsevolod V. Gurevich, and T. M. Iverson

7. Detection of Structural Waters and Their Role in Structural Dynamics of Rhodopsin Activation
Liwen Wang and Mark Chance

8. Probing Conformational Changes in Rhodopsin Using Hydrogen-Deuterium Exchange Coupled to Mass Spectrometry
Tivadar Orban and Yaroslav Tsybovsky

9. Analysis of Conformational Changes in Rhodopsin by Histidine Hydrogen-Deuterium Exchange
David T. Lodowski and Masaru Miyagi

10. Investigation of Rhodopsin Dynamics in its Signaling State by Solid-State Deuterium NMR Spectroscopy
Andrey V. Struts, Udeep Chawla, Suchithranga M.D.C. Perera, and Michael F. Brown

11. Sequential Structural Changes in Rhodopsin Occurring upon Photoactivation
Naoki Kimata, Andreyah Pope, Dawood Rashid, Philip J. Reeves, and Steven O. Smith

12. Dynamic Single-Molecule Force Spectroscopy of Rhodopsin in Native Membranes
Paul S.-H. Park and Daniel J. Müller

Part IV: Rhodopsin Supramolecular Organization and its Complexes with Partner Proteins

13. High-Resolution Atomic Force Microscopy Imaging of Rhodopsin in Rod Outer Segment Disk Membranes
Patrick D. Bosshart, Andreas Engel, and Dimitrios Fotiadis

14. Detection of Rhodopsin Dimerization in situ by PIE-FCCS, a Time-Resolved Fluorescence Spectroscopy
Adam W. Smith

15. Oligomeric State of Rhodopsin within Rhodopsin-Transducin Complex Probed with Succinylated Concanavalin A
Beata Jastrzebska

16. Quantification of Arrestin-Rhodopsin Binding Stoichiometry
Ciara C.M. Lally and Martha E. Sommer

17. Rhodopsin Transient Complexes Investigated by Systems Biology Approaches
Daniele Dell'Orco

Part V: Rhodopsin and Photoreceptors

18. Three Dimensional Architecture of Murine Rod Cilium Revealed by Cryo-EM
Theodore G. Wensel and Jared C. Gilliam

19. Monitoring of Rhodopsin Trafficking and Mistrafficking in Live Photoreceptors
Kerrie H. Lodowski and Yoshikazu Imanishi

20. Measurements of Rhodopsin Diffusion within Signaling Membrane Microcompartments in Live Photoreceptors
Mehdi Najafi and Peter D. Calvert

Part VI: Treatment Strategies of Retinal Degenerative Disease

21. Kinetics of Rhodopsin's Chromophore Monitored in a Single Photoreceptor
Leopold Adler IV, Nicholas P. Boyer, Chunhe Chen, and Yiannis Koutalos

22. Supplementation with Vitamin A Derivatives to Rescue Vision in Animal Models of Degenerative Retinal Diseases
Lindsay Perusek, Akiko Maeda, and Tadao Maeda

23. Sustained Delivery of Retinoids to Prevent Photoreceptor Death
Peter H. Tang and Rosalie K. Crouch

24. High-Throughput Screening Assays to Identify Small-Molecules Preventing Photoreceptor Degeneration Caused by the Rhodopsin P23H Mutation
Yuanyuan Chen and Hong Tang

25. Gene Therapy to Rescue Retinal Degeneration Caused by Mutations in Rhodopsin
Brian P. Rossmiller, Renee C. Ryals, and Alfred S. Lewin