GO:0102354 11-cis-retinol dehydrogenase (NADP+) activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0102354 defines the NADP+-dependent oxidation of 11-cis-retinol to 11-cis-retinal, a key step in the visual cycle.
• RDH5 and RDH10 are the principal enzymes experimentally shown to carry this activity in the retinal pigment epithelium [1,2].
• The reaction is reversible and uses NADP+ as the preferred cofactor, distinguishing it from NAD+-dependent retinol dehydrogenases [1,4].
• Loss of 11-cis-retinol dehydrogenase activity is linked to fundus albipunctatus, a hereditary retinal disorder.
• The enzyme interacts with visual cycle proteins and the opsin chromophore, influencing photoreceptor function [1,3].
• CRISPR knockout, point-mutation, and knock-in models enable precise dissection of this activity in retinal cells [1,2].
Description
11-cis-retinol dehydrogenase (NADP+) activity, encoded by the Gene Ontology term GO:0102354, catalyzes the reversible oxidation of 11-cis-retinol to 11-cis-retinal with concomitant reduction of NADP+ to NADPH. This reaction is a critical node in the visual cycle, the biochemical pathway that regenerates the chromophore required for light detection in photoreceptors [1,2]. The enzyme belongs to the short-chain alcohol dehydrogenase family and is highly expressed in the retinal pigment epithelium (RPE). Researchers study this activity to understand how the retina maintains visual pigment regeneration and how defects contribute to inherited retinal degenerations [2,3]. The term is also relevant to broader retinoid metabolism because 11-cis-retinal serves as a precursor for visual pigments and can influence retinoic acid signaling. Experimental models, including Rdh5 knockout mice, have been instrumental in characterizing the enzymatic activity and its physiological role. The NADP+ dependence of this dehydrogenase distinguishes it from other retinol dehydrogenases that prefer NAD+ [1,4].
11-cis-retinol dehydrogenase (NADP+) activity At A Glance
| GO ID | GO:0102354 |
|---|---|
| GO term | 11-cis-retinol dehydrogenase (NADP+) activity |
| Ontology | molecular_function |
| Synonym | 11-cis-retinol dehydrogenase activity |
| Major function | Catalyzes the NADP+-dependent oxidation of 11-cis-retinol to 11-cis-retinal |
| Reaction | 11-cis-retinol + NADP+ = 11-cis-retinal + NADPH + H+ |
| Cofactor | NADP+ (preferred over NAD+) |
| Localization | Retinal pigment epithelium (RPE) and other retinoid-metabolizing tissues |
| Related genes | RDH5, RDH10, and other short-chain dehydrogenases |
What Is GO:0102354?
GO:0102354 describes the catalysis of the reaction: 11-cis-retinol + NADP+ = 11-cis-retinal + NADPH + H+. In other words, it is the NADP+-dependent oxidation of 11-cis-retinol to 11-cis-retinal, a reversible step in the visual cycle.
Why Is 11-cis-retinol dehydrogenase (NADP+) activity Important in Cell Biology?
This activity is essential for regenerating the visual chromophore 11-cis-retinal, which is required for photoreceptor function and vision [1,2]. Defects in the enzyme lead to impaired dark adaptation and retinal degeneration, as seen in fundus albipunctatus. Understanding its mechanism provides insights into retinoid metabolism and potential therapeutic targets for retinal diseases [3,5].
• Critical for the visual cycle and photoreceptor health.
• Mutations in RDH5 cause fundus albipunctatus, a retinal disorder.
• Interacts with visual cycle proteins and opsin [1,3].
• NADP+ preference distinguishes it from other retinol dehydrogenases.
• Contributes to retinoic acid homeostasis.
• Target for gene therapy and pharmacological modulation.
• Model for studying short-chain dehydrogenase mechanisms.
• Relevant to age-related macular degeneration and retinitis pigmentosa.
Molecular Mechanism of 11-cis-retinol dehydrogenase (NADP+) activity
Substrate binding and cofactor preference
In simple terms: The enzyme grabs 11-cis-retinol and NADP+ to start the reaction.
The enzyme binds 11-cis-retinol and NADP+ in a sequential manner. RDH5 and RDH10 exhibit a preference for NADP+ over NAD+, as demonstrated by enzymatic assays [1,4]. The binding site accommodates the cis configuration of the retinol substrate, ensuring specificity.
Catalytic oxidation and product release
In simple terms: The enzyme removes hydrogens from retinol to make retinal.
Oxidation of 11-cis-retinol involves hydride transfer to NADP+, forming 11-cis-retinal and NADPH. The reaction is reversible, and the equilibrium can favor either direction depending on substrate and cofactor concentrations. Product release is facilitated by conformational changes in the active site.
Interaction with visual cycle proteins
In simple terms: The enzyme works together with other proteins to recycle the visual pigment.
RDH10 interacts with visual cycle proteins such as RPE65 and CRALBP, which channel retinoids between enzymatic steps. These interactions enhance the efficiency of 11-cis-retinal production and prevent toxic accumulation of retinoids [1,3].
Regulation by retinoid availability and cellular redox state
In simple terms: The enzyme's speed depends on how much retinol and NADP+ are available.
Enzyme activity is modulated by the availability of 11-cis-retinol and the NADP+/NADPH ratio. High NADPH levels can inhibit the oxidative direction, while NADP+ excess promotes it [1,4]. This redox sensitivity links the enzyme to cellular metabolic status.
Key Genes Involved in GO:0102354 11-cis-retinol dehydrogenase (NADP+) activity
The following genes and proteins are directly implicated in 11-cis-retinol dehydrogenase (NADP+) activity or its regulation.
| Gene | Major Role | Research Relevance |
|---|---|---|
| RDH5 | Primary 11-cis-retinol dehydrogenase in RPE | Mutations cause fundus albipunctatus; knockout models available |
| RDH10 | 11-cis-retinol dehydrogenase with broad expression | Interacts with visual cycle proteins; knockout embryonic lethal |
| RPE65 | Retinoid isomerase in visual cycle | Interacts with RDH10; mutations cause Leber congenital amaurosis |
| CRALBP | Retinaldehyde-binding protein | Channels retinoids to RDH; mutations cause retinal dystrophy |
| LRAT | Lecithin retinol acyltransferase | Produces retinyl esters for visual cycle |
| RGR | Retinal G protein-coupled receptor | Binds 11-cis-retinal; interacts with RDH5 |
| OPSIN | Photoreceptor pigment protein | Binds 11-cis-retinal; interaction with RDH5 studied |
| RDH11 | Retinol dehydrogenase | May compensate for RDH5 loss |
| RDH12 | Retinal dehydrogenase | Mutations cause retinal dystrophy; related activity |
| ALDH1A1 | Retinaldehyde dehydrogenase | Produces retinoic acid from retinal |
| ALDH1A2 | Retinaldehyde dehydrogenase | Retinoic acid synthesis |
| CYP26A1 | Retinoic acid hydroxylase | Degrades retinoic acid; affects retinoid balance |
| CRBP1 | Cellular retinol-binding protein | Binds retinol; influences substrate availability |
| CRABP1 | Cellular retinoic acid-binding protein | Modulates retinoic acid signaling |
| RDH13 | Retinol dehydrogenase | Potential alternative activity |
| RDH14 | Retinol dehydrogenase | Expressed in retina; function under study |
| SDR16C5 | Short-chain dehydrogenase | Related to retinol metabolism |
| SDR16C6 | Short-chain dehydrogenase | Related to retinol metabolism |
How Is 11-cis-retinol dehydrogenase (NADP+) activity Regulated?
The activity of 11-cis-retinol dehydrogenase is regulated by substrate availability, cofactor ratios, and protein-protein interactions within the visual cycle. NADP+/NADPH balance directly influences the direction of the reaction. RPE65 and CRALBP facilitate substrate channeling, enhancing enzymatic efficiency. Retinoic acid levels can feedback on retinoid metabolism through enzymes such as CYP26A1.
11-cis-retinol dehydrogenase (NADP+) activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| RDH5 | Fundus albipunctatus | Rdh5 knockout mouse; patient-derived iPSC-RPE |
| RDH10 | Retinoic acid deficiency, embryonic lethality | Conditional knockout mouse |
| RDH12 | Retinal dystrophy | Knock-in mouse with patient mutation |
| ALDH1A1 | Cancer, retinoic acid signaling | Overexpression cell lines |
| CYP26A1 | Retinoic acid catabolism disorders | Knockout zebrafish |
Fundus albipunctatus
Mutations in RDH5, which encodes the primary 11-cis-retinol dehydrogenase, cause fundus albipunctatus, a rare retinal disorder characterized by night blindness and white dots in the fundus. Knockout mouse models of Rdh5 show delayed dark adaptation and reduced 11-cis-retinal regeneration.
Retinal degenerations
Impaired 11-cis-retinol dehydrogenase activity can lead to accumulation of toxic retinoids and photoreceptor degeneration. RDH12 mutations are associated with severe early-onset retinal dystrophy, highlighting the importance of retinol dehydrogenases in retinal health.
Retinoic acid signaling disorders
Altered 11-cis-retinol dehydrogenase activity affects retinoic acid homeostasis, which is critical for development and differentiation. Dysregulation of retinoic acid signaling is implicated in various cancers and developmental defects.
From 11-cis-retinol dehydrogenase (NADP+) activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does RDH5 loss impair visual cycle? | Rdh5 knockout mouse |
| How does RDH10 interact with RPE65? | RDH10 knockout or knock-in tagged RDH10 |
| What is the effect of point mutations in RDH5? | CRISPR point-mutation knock-in in RPE cells |
| Can overexpression rescue activity? | RDH5 overexpression in RPE cells |
| What is the role of NADP+ binding? | Site-directed mutagenesis of cofactor-binding residues |
| How does 11-cis-retinal interact with opsin? | In vitro binding assays with purified proteins |
How to Study the 11-cis-retinol dehydrogenase (NADP+) activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| NADPH absorbance assay | Enzyme activity | Kinetic studies of RDH5/RDH10 |
| HPLC | Retinoid levels | Visual cycle analysis in knockout mice |
| Co-immunoprecipitation | Protein-protein interactions | Visual cycle complex assembly |
| CRISPR knockout screen | Gene essentiality | Identifying modifiers of visual cycle |
| Site-directed mutagenesis | Cofactor specificity | Mapping NADP+ binding site |
| Immunofluorescence | Protein localization | RPE cell polarity studies |
| RNA-seq | Gene expression changes | Retinoid metabolism profiling |
| Mass spectrometry | Protein identification | Purification of dehydrogenase complexes |
Enzymatic activity assays
In vitro assays using retinal membrane fractions or purified recombinant enzymes measure the conversion of 11-cis-retinol to 11-cis-retinal by monitoring NADPH production at 340 nm [1,4].
Retinoid profiling by HPLC
High-performance liquid chromatography (HPLC) quantifies retinoid species in tissues or cell cultures, allowing assessment of 11-cis-retinal and 11-cis-retinol levels.
Protein interaction studies
Co-immunoprecipitation and pull-down assays identify interactions between RDH10 and visual cycle proteins such as RPE65 and CRALBP.
CRISPR-based genetic screens
Genome-wide CRISPR knockout screens in RPE cells can identify genes that modify 11-cis-retinol dehydrogenase activity and visual cycle function.
How CRISPR Can Be Used to Study GO:0102354 11-cis-retinol dehydrogenase (NADP+) activity
Knockout
CRISPR knockout of RDH5 or RDH10 in RPE cells or mouse models abolishes 11-cis-retinol dehydrogenase activity, leading to impaired visual cycle and retinal degeneration. These models are used to study the contribution of each enzyme to 11-cis-retinal production.
Point Mutation
Introducing patient-specific point mutations in RDH5 (e.g., those found in fundus albipunctatus) via CRISPR base editing or homology-directed repair allows assessment of enzymatic activity and protein stability.
Knock-in
Knock-in of tagged RDH10 or RDH5 (e.g., GFP or HA tags) enables live-cell imaging and proteomic analysis of the enzyme in its native context.
Overexpression
Overexpression of RDH5 or RDH10 in cell lines can rescue loss-of-function phenotypes and provide sufficient material for biochemical purification and structural studies.
How EDITGENE Supports 11-cis-retinol dehydrogenase (NADP+) activity Research
Researchers studying 11-cis-retinol dehydrogenase (NADP+) activity-related genes often need to determine whether a candidate gene is causally involved in visual cycle regulation or retinal disease. EDITGENE provides CRISPR-based cell models and screening services to accelerate this discovery process.
Contact EDITGENE today to design your custom CRISPR model for 11-cis-retinol dehydrogenase (NADP+) activity research.
Frequently Asked Questions About 11-cis-retinol dehydrogenase (NADP+) activity
What is 11-cis-retinol dehydrogenase (NADP+) activity?
It is the enzyme activity that converts 11-cis-retinol to 11-cis-retinal using NADP+ as a cofactor, encoded by GO:0102354.
What genes are involved in 11-cis-retinol dehydrogenase (NADP+) activity?
RDH5 and RDH10 are the primary genes, with additional contributions from RDH11, RDH12, and visual cycle proteins [1,2].
What diseases are associated with 11-cis-retinol dehydrogenase deficiency?
Mutations in RDH5 cause fundus albipunctatus, and RDH12 mutations lead to severe retinal dystrophy.
How is 11-cis-retinol dehydrogenase activity measured?
It is measured by enzymatic assays monitoring NADPH production or by HPLC quantification of retinoids [1,4].
What is the role of NADP+ in this reaction?
NADP+ acts as the electron acceptor, being reduced to NADPH during the oxidation of 11-cis-retinol.
Can CRISPR be used to study 11-cis-retinol dehydrogenase?
Yes, CRISPR knockout and knock-in models in RPE cells and mice are used to dissect gene function.
What is the difference between RDH5 and RDH10?
RDH5 is RPE-specific and primarily responsible for 11-cis-retinol oxidation, while RDH10 is more broadly expressed and interacts with visual cycle proteins [1,2].
What are the symptoms of fundus albipunctatus?
Night blindness and delayed dark adaptation, often with white dots in the retina.
Is 11-cis-retinol dehydrogenase activity reversible?
Yes, the reaction is reversible, and the direction depends on substrate and cofactor concentrations.
How does 11-cis-retinol dehydrogenase interact with opsin?
It provides 11-cis-retinal, which binds opsin to form the visual pigment rhodopsin.
Conclusion
11-cis-retinol dehydrogenase (NADP+) activity (GO:0102354) is a cornerstone of the visual cycle, catalyzing the NADP+-dependent oxidation of 11-cis-retinol to 11-cis-retinal. Its dysfunction leads to retinal diseases such as fundus albipunctatus, making it a key target for gene therapy and pharmacological intervention. CRISPR-based models and biochemical assays continue to unravel its mechanism and regulation.
References
- 1. Farjo KM et al.. 2009. The 11-cis-retinol dehydrogenase activity of RDH10 and its interaction with visual cycle proteins.. Invest Ophthalmol Vis Sci 50(11):5089-97 PMID: 19458327
- 2. Jang GF et al.. 2001. Characterization of a dehydrogenase activity responsible for oxidation of 11-cis-retinol in the retinal pigment epithelium of mice with a disrupted RDH5 gene. A model for the human hereditary disease fundus albipunctatus.. J Biol Chem 276(35):32456-65 PMID: 11418621
- 3. Chen P et al.. 2001. Interaction of 11-cis-retinol dehydrogenase with the chromophore of retinal g protein-coupled receptor opsin.. J Biol Chem 276(24):21098-104 PMID: 11274198
- 4. Simon A et al.. 1995. The retinal pigment epithelial-specific 11-cis retinol dehydrogenase belongs to the family of short chain alcohol dehydrogenases.. J Biol Chem 270(3):1107-12 PMID: 7836368
- 5. Napoli JL et al.. 1995. Enzymes and binding proteins affecting retinoic acid concentrations.. J Steroid Biochem Mol Biol 53(1-6):497-502 PMID: 7626500
