ArticleInvestigative ophthalmology & visual science2015
A High-Throughput Drug Screening Strategy for Detecting Rhodopsin P23H Mutant Rescue and Degradation.
Article in Investigative ophthalmology & visual science, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 18 citations in OpenAlex.
- Nonretinoid chaperones improve rhodopsin homeostasis in a mouse model of retinitis pigmentosa.JCI insight · 2022Article
- Pharmacological clearance of misfolded rhodopsin for the treatment of RHO-associated retinitis pigmentosa.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2020Article
- Screening of Chemical Libraries Using a Yeast Model of Retinal Disease.SLAS discovery : advancing life sciences R & D · 2019Article
- Rhodopsin Oligomerization and Aggregation.The Journal of membrane biology · 2019Review
- Stereospecific modulation of dimeric rhodopsin.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2019Article
- Müller glia phagocytose dead photoreceptor cells in a mouse model of retinal degenerative disease.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2019Article
- A Rhodopsin Transport Assay by High-Content Imaging Analysis.Journal of visualized experiments : JoVE · 2019Article
- Comparison of the molecular properties of retinitis pigmentosa P23H and N15S amino acid replacements in rhodopsin.PloS one · 2019Article
- Misfolded rhodopsin mutants display variable aggregation properties.Biochimica et biophysica acta. Molecular basis of disease · 2018Article
- A novel small molecule chaperone of rod opsin and its potential therapy for retinal degeneration.Nature communications · 2018Article
- Ligand channel in pharmacologically stabilized rhodopsin.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Rescue of mutant rhodopsin traffic by metformin-induced AMPK activation accelerates photoreceptor degeneration.Human molecular genetics · 2017Article
- Transcriptome profiling of NIH3T3 cell lines expressing opsin and the P23H opsin mutant identifies candidate drugs for the treatment of retinitis pigmentosa.Pharmacological research · 2017Article
- Wild-type opsin does not aggregate with a misfolded opsin mutant.Biochimica et biophysica acta · 2016Article
- A small molecule mitigates hearing loss in a mouse model of Usher syndrome III.Nature chemical biology · 2016Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
Abstract
purposeInherent instability of the P23H mutant opsin accounts for approximately 10% of autosomal dominant retinitis pigmentosa cases. Our purpose was to develop an overall set of reliable screening strategies to assess if either stabilization or enhanced degradation of mutant rhodopsin could rescue rod photoreceptors expressing this mutant protein. These strategies promise to reveal active compounds and clarify molecular mechanisms of biologically important processes, such as inhibition of target degradation or enhanced target folding.
methodsCell-based bioluminescence reporter assays were developed and validated for high-throughput screening (HTS) of compounds that promote either stabilization or degradation of P23H mutant opsin. Such assays were further complemented by immunoblotting and image-based analyses.
resultsTwo stabilization assays of P23H mutant opsin were developed and validated, one based on β-galactosidase complementarity and a second assay involving bioluminescence resonance energy transfer (BRET) technology. Moreover, two additional assays evaluating mutant protein degradation also were employed, one based on the disappearance of luminescence and another employing the ALPHA immunoassay. Imaging of cells revealed the cellular localization of mutant rhodopsin, whereas immunoblots identified changes in the aggregation and glycosylation of P23H mutant opsin.
conclusionsOur findings indicate that these initial HTS and following assays can identify active therapeutic compounds, even for difficult targets such as mutant rhodopsin. The assays are readily scalable and their function has been proven with model compounds. High-throughput screening, supported by automated imaging and classic immunoassays, can further characterize multiple steps and pathways in the biosynthesis and degradation of this essential visual system protein.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.