ArticleThe Journal of biological chemistry2012
Structural tuning of the fluorescent protein iLOV for improved photostability.
Article in The Journal of biological chemistry, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 65 papers.
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Who cites it
65 citing papers in PubMed, 169 citations in OpenAlex.
- Optically detected and radio wave-controlled spin chemistry in flavoproteins.Nature biotechnology · 2026Article
- Protein Electrostatics Tune the Singlet-Triplet Energy Gap in Natural and Engineered Phototropin Light-Oxygen-Voltage (LOV) Domains.Journal of the American Chemical Society · 2026Article
- Insertion of Fluorescent Proteins Near the Plug Domain of MotB Generates Functional Stator Complexes.MicrobiologyOpen · 2025Article
- Construction of enterovirus G expressing reporter genes for antiviral drug screening assays.BMC veterinary research · 2025Article
- Article
- Electronic Structure Methods for Simulating Flavin's Spectroscopy and Photophysics: Comparison of Multi-reference, TD-DFT, and Single-Reference Wave Function Methods.The journal of physical chemistry. B · 2024Article
- Rational Design of a Circularly Permuted Flavin-Based Fluorescent Protein.Chembiochem : a European journal of chemical biology · 2024Article
- Light Control in Microbial Systems.International journal of molecular sciences · 2024Review
- Genetically Encoded Biosensors for the Fluorescence Detection of OSensors (Basel, Switzerland) · 2023Review
- Deep Mutational Scanning of an Oxygen-Independent Fluorescent Protein CreiLOV for Comprehensive Profiling of Mutational and Epistatic Effects.ACS synthetic biology · 2023Article
- Alternative Strategy for Spectral Tuning of Flavin-Binding Fluorescent Proteins.The journal of physical chemistry. B · 2023Article
- Development and Characterization of Flavin-Binding Fluorescent Proteins, Part II: Advanced Characterization.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Anaerobic fluorescent reporters for live imaging ofFrontiers in microbiology · 2023Article
- Experimental Characterization ofACS omega · 2022Article
- Agrobacterium expressing a type III secretion system delivers Pseudomonas effectors into plant cells to enhance transformation.Nature communications · 2022Article
- Rapid directed molecular evolution of fluorescent proteins in mammalian cells.Protein science : a publication of the Protein Society · 2022Article
- Practical observations on the use of fluorescent reporter systems in Clostridioides difficile.Antonie van Leeuwenhoek · 2022Article
- Enhanced small green fluorescent proteins as a multisensing platform for biosensor development.Frontiers in bioengineering and biotechnology · 2022Article
- Peripheral Methionine Residues Impact Flavin Photoreduction and Protonation in an Engineered LOV Domain Light Sensor.Biochemistry · 2021Article
- Computational Investigation of Structural and Spectroscopic Properties of LOV-Based Proteins with Improved Fluorescence.The journal of physical chemistry. B · 2021Article
5 more citing papers are in PubMed but not listed here.
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Authors and funding
8 authors at 3 institutions in 3 countries.
Funding
Abstract
Fluorescent proteins derived from light, oxygen, or voltage (LOV) domains offer advantages over green fluorescent protein (GFP) from their small size and efficacy under anaerobic conditions. The flavoprotein improved LOV (iLOV) was engineered from the blue light receptor phototropin as a reporter of viral infection. To inform the molecular basis for the improved, photoreversible, fluorescent properties of iLOV, we employed directed evolution and determined five LOV crystallographic structures. Comparative structural analyses between iLOV and its progenitors reveal mutation-induced constraints in the environment of the flavin mononucleotide (FMN) chromophore; in iLOV, the methyl group of Thr-394 "crowds" the FMN isoalloxazine ring, Leu-470 triggers side chain "flipping" of Leu-472, and the terminal FMN phosphate shows increased anchoring. We further engineered iLOV variants that are readily detectable in bacterial and mammalian cells due to order-of-magnitude photostability increases. Structure determination of a resulting representative photostable iLOV (phiLOV) variant reveals additional constraints on the chromophore. Aromatic residues Tyr-401 and Phe-485 in phiLOV sandwich the FMN isoalloxazine ring from both sides, whereas Ser-390 anchors the side chain of FMN-interacting Gln-489 Our combined structural and mutational results reveal that constraining the FMN fluorophore yields improved photochemical properties for iLOV and its new photostable derivative. These findings provide a framework for structural fine-tuning of LOV scaffold proteins to maximize their potential as oxygen-independent fluorescent reporters.
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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.