ArticleProceedings of the National Academy of Sciences of the United States of America2014
Computational design of a red fluorophore ligase for site-specific protein labeling in living cells.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 71 citations in OpenAlex.
- A globular protein exhibits rare phase behavior and forms chemically regulated orthogonal condensates in cells.Nature communications · 2025Article
- Lipoic acid attachment to proteins: stimulating new developments.Microbiology and molecular biology reviews : MMBR · 2024Review
- PocketOptimizer 2.0: A modular framework for computer-aided ligand-binding design.Protein science : a publication of the Protein Society · 2023Article
- Rational design of enzyme activity and enantioselectivity.Frontiers in bioengineering and biotechnology · 2023Review
- Understanding and Engineering Glycine Cleavage System and Related Metabolic Pathways for C1-Based Biosynthesis.Advances in biochemical engineering/biotechnology · 2022Review
- Coumarin luciferins and mutant luciferases for robust multi-component bioluminescence imaging.Chemical science · 2021Article
- Strategies for Site-Specific Labeling of Receptor Proteins on the Surfaces of Living Cells by Using Genetically Encoded Peptide Tags.Chembiochem : a European journal of chemical biology · 2021Review
- NanoFAST: structure-based design of a small fluorogen-activating protein with only 98 amino acids.Chemical science · 2021Article
- Multicomponent Bioluminescence Imaging with a π-Extended Luciferin.Journal of the American Chemical Society · 2020Article
- Synthetic biology in various cellular and molecular fields: applications, limitations, and perspective.Molecular biology reports · 2020Review
- Engineering of a Red Fluorogenic Protein/Merocyanine Complex for Live-Cell Imaging.Chembiochem : a European journal of chemical biology · 2020Article
- Addition of Isocyanide-Containing Amino Acids to the Genetic Code for Protein Labeling and Activation.ACS chemical biology · 2019Article
- Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy.Bio-protocol · 2019Article
- Computational Aminoacyl-tRNA Synthetase Library Design for Photocaged Tyrosine.International journal of molecular sciences · 2019Article
- Control of Intra- versus Extracellular Bioorthogonal Catalysis Using Surface-Engineered Nanozymes.ACS nano · 2019Article
- Sequence-based prediction of permissive stretches for internal protein tagging and knockdown.BMC biology · 2017Article
- Protein labeling for live cell fluorescence microscopy with a highly photostable renewable signal.Chemical science · 2017Article
- Crystal structure of lipoate-bound lipoate ligase 1, LipL1, from Plasmodium falciparum.Proteins · 2017Article
- Breaking the color barrier - a multi-selective antibody reporter offers innovative strategies of fluorescence detection.Journal of cell science · 2017Article
- Broad substrate tolerance of tubulin tyrosine ligase enables one-step site-specific enzymatic protein labeling.Chemical science · 2017Article
Corrections and comments
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Authors and funding
13 authors at 5 institutions in 2 countries.
Funding
Abstract
Chemical fluorophores offer tremendous size and photophysical advantages over fluorescent proteins but are much more challenging to target to specific cellular proteins. Here, we used Rosetta-based computation to design a fluorophore ligase that accepts the red dye resorufin, starting from Escherichia coli lipoic acid ligase. X-ray crystallography showed that the design closely matched the experimental structure. Resorufin ligase catalyzed the site-specific and covalent attachment of resorufin to various cellular proteins genetically fused to a 13-aa recognition peptide in multiple mammalian cell lines and in primary cultured neurons. We used resorufin ligase to perform superresolution imaging of the intermediate filament protein vimentin by stimulated emission depletion and electron microscopies. This work illustrates the power of Rosetta for major redesign of enzyme specificity and introduces a tool for minimally invasive, highly specific imaging of cellular proteins by both conventional and superresolution microscopies.
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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.