What site-directed labeling studies tell us about the mechanism of rhodopsin activation and G-protein binding
Open accessAbstract
Rhodopsin is the photoreceptor protein responsible for dim-light vision in mammals. Due to extensive biophysical, structural and computational analysis of this membrane protein, it is presently the best understood G-protein coupled receptor (GPCR). Here I briefly review one approach that has been extensively used to identify dynamic and structural changes in rhodopsin--the use of site-directed labeling methods (SDL) coupled with electron paramagnetic resonance (EPR) and fluorescence spectroscopy. These SDL studies involve introducing individual cysteine residues into the receptor, then labeling them with cysteine-reactive probes for subsequent analysis by the appropriate spectroscopy. I will give a brief overview of how SDL methods are carried out and how the data is analyzed. Then, I will discuss how SDL studies were carried out on rhodopsin, and how they were used to identify a key structural change that occurs in rhodopsin upon activation--movement of transmembrane helix 6 (TM6). I will also briefly discuss how the SDL studies of rhodopsin compare with SDL studies of other GPCRs, and compare the SDL data with early and recent crystal structures of rhodopsin. Finally, I will discuss why the TM6 movement is required for rhodopsin to couple with the G-protein transducin, and speculate how this mechanism might be a universal method used by all GPCRs to bind G-proteins and perhaps other proteins involved in visual signal transduction, such as arrestin and rhodopsin kinase.
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Rhodopsin is the main protein of ROS disk and plasma membranes, making up 90% and 75% of their protein content respectively.
“Rhodopsin is the main component of disk and plasma membranes of ROS, accounting for 90 and 75% of their protein content.”
From Chemistry of the Retinoid (Visual) Cycle (2014) · cites this paper as [321]
The Abstract identifies rhodopsin as a photoreceptor membrane protein, while the ‘Helix 6 in rhodopsin moves upon light activation’ section discusses structural changes rather than ROS membrane composition (page numbers unavailable). Searches for disk and plasma membranes and for the 90% and 75% figures found no passage stating rhodopsin’s share of their protein content. (LLM, conf=0.76)
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Chemistry of the Retinoid (Visual) Cycle
Kiser, P. D., Golczak, M., & Palczewski, K. (2014) · Chemical Reviews · doi:10.1021/cr400107q · Cited by 353 · see citing works
- Authors
- Farrens, D. L.
- Affiliations
- Oregon Health & Science University (US)
- Corresponding author
- Farrens, David L.
- ORCID
- Farrens, David L.
- Venue
- Photochemical & Photobiological Sciences
- Published
- 2010-11
- Volume
- 9
- Issue
- 11
- Pages
- 1466-1474
- ISSN
- 1474-905X (ISSN-L)
- 1474-9092
- PMC
- PMC3901699
- OpenAlex
- W2021192083
- Cited by
- 28
- References
- 69
- Citations by year
- 2026: 1 · 2022: 2 · 2020: 2 · 2019: 1 · 2017: 1 · 2016: 2 · 2015: 2 · 2014: 3 · 2013: 6 · 2012: 5
- FWCI
- 1.81
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Photochemical & Photobiological Sciences (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health
- National Institute on Drug Abuse (DA018169, R01DA018169)
- National Eye Institute (EY015436, R01EY015436)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Photoreceptor and optogenetics research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Electron Spin Resonance Studies
- Keywords
- rhodopsin, G protein-coupled receptors, G protein activation, transducin, transmembrane helix 6, site-specific labeling, electron spin resonance, fluorescence spectroscopy, arrestin, rhodopsin kinase, phototransduction, GPCR crystal structures
- MeSH terms
- Animals, Electron Spin Resonance Spectroscopy, Humans, Models, Molecular, Protein Binding, Protein Conformation, Rhodopsin, Spectrometry, Fluorescence, GTP-Binding Proteins
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