ArticleProceedings of the National Academy of Sciences of the United States of America2010
A fluorophore ligase for site-specific protein labeling inside living cells.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 124 papers.
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Who cites it
124 citing papers in PubMed, 287 citations in OpenAlex.
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- Library-on-Library Intercellular Labeling for Selection of Biotin Ligase and Acceptor Peptides.Chembiochem : a European journal of chemical biology · 2026Article
- Synthetic fluorophores for live-cell fluorescence microscopy and biosensing.Nature chemical biology · 2025Review
- Homogeneous antibody-drug conjugates with dual payloads: potential, methods and considerations.mAbs · 2025Review
- Lipoic acid attachment to proteins: stimulating new developments.Microbiology and molecular biology reviews : MMBR · 2024Review
- Chemical tags and beyond: Live-cell protein labeling technologies for modern optical imaging.Smart molecules : open access · 2023Review
- Fluorogenic Cyclopropenones for Multicomponent, Real-Time Imaging.Journal of the American Chemical Society · 2022Article
- Molecular probes for cellular imaging of post-translational proteoforms.RSC chemical biology · 2022Review
- Diverse protein manipulations with genetically encoded glutamic acid benzyl ester.Chemical science · 2021Article
- Enzymatic Methods for the Site-Specific Radiolabeling of Targeting Proteins.Molecules (Basel, Switzerland) · 2021Review
- 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
- A Genetic Code Expansion-Derived Molecular Beacon for the Detection of Intracellular Amyloid-β Peptide Generation.Angewandte Chemie (Weinheim an der Bergstrasse, Germany) · 2021Article
- A Genetic Code Expansion-Derived Molecular Beacon for the Detection of Intracellular Amyloid-β Peptide Generation.Angewandte Chemie (International ed. in English) · 2021Article
- A Bioorthogonal Click Chemistry Toolbox for Targeted Synthesis of Branched and Well-Defined Protein-Protein Conjugates.Angewandte Chemie (International ed. in English) · 2020Article
- Topological Features of Histone H2A Monoubiquitination.Doklady. Biochemistry and biophysics · 2020Article
- Engineering of a Red Fluorogenic Protein/Merocyanine Complex for Live-Cell Imaging.Chembiochem : a European journal of chemical biology · 2020Article
- Single-Virus Tracking: From Imaging Methodologies to Virological Applications.Chemical reviews · 2020Review
64 more citing papers are in PubMed but not listed here.
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
Abstract
Biological microscopy would benefit from smaller alternatives to green fluorescent protein for imaging specific proteins in living cells. Here we introduce PRIME (PRobe Incorporation Mediated by Enzymes), a method for fluorescent labeling of peptide-fused recombinant proteins in living cells with high specificity. PRIME uses an engineered fluorophore ligase, which is derived from the natural Escherichia coli enzyme lipoic acid ligase (LplA). Through structure-guided mutagenesis, we created a mutant ligase capable of recognizing a 7-hydroxycoumarin substrate and catalyzing its covalent conjugation to a transposable 13-amino acid peptide called LAP (LplA Acceptor Peptide). We showed that this fluorophore ligation occurs in cells in 10 min and that it is highly specific for LAP fusion proteins over all endogenous mammalian proteins. By genetically targeting the PRIME ligase to specific subcellular compartments, we were able to selectively label spatially distinct subsets of proteins, such as the surface pool of neurexin and the nuclear pool of actin.
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Registered trials
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.