ArticleEnvironmental microbiology2014
In situ visualization of newly synthesized proteins in environmental microbes using amino acid tagging and click chemistry.
Article in Environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 125 papers.
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Who cites it
125 citing papers in PubMed, 245 citations in OpenAlex.
- Elucidating the role of the gut microbiota in the physiological effects of dietary fiber.Microbiome · 2022Trial
- From dividing to dormant: embracing the full activity spectrum for environmental microorganisms.Microbiology and molecular biology reviews : MMBR · 2026Review
- Advancements and applications of click chemistry in protein labeling and bioconjugation.RSC advances · 2026Review
- Mass spectrometry proteomics for studying mitostasis.Protein science : a publication of the Protein Society · 2026Review
- Dual-BONCAT reveals distinct subpopulations of anabolically active cells.Applied and environmental microbiology · 2026Article
- Rainfall-induced microbial resuscitation reveals functional decoupling across biocrust succession.The ISME journal · 2026Article
- Deep-sea anaerobic microbial communities couple degradation of insoluble chitin to extracellular electron transfer.The ISME journal · 2026Article
- Tracking active heterotrophic microbial communities in the Guaymas Basin deep biosphere with BONCAT-FACS.ISME communications · 2026Article
- From Molecule to Meaning: Click and Bioorthogonal Chemical Reporters for Plant Systems, Biological Imaging, and Artistic Expression.Chembiochem : a European journal of chemical biology · 2026Review
- SIESTA Project: Svalbard summer 2025 expedition report.Open research Europe · 2026Article
- Single-cell heterotrophic activity in deep-ocean prokaryotic communities quantified by BONCAT and microautoradiography.ISME communications · 2026Article
- Illuminating the newly produced viruses within the virosphere with bioorthogonal noncanonical amino acid tagging and single-virus genomic sequencing technologies.ISME communications · 2026Article
- Lactulose selectively stimulates members of the gut microbiota, as determined by multi-modal activity-based sorting.Gut microbes · 2025Article
- Article
- Activity-targeted metaproteomics uncovers rare syntrophic bacteria central to anaerobic community metabolism.Nature microbiology · 2025Article
- Using BONCAT to dissect the proteome ofmSphere · 2025Article
- The activity of soil microbial taxa in the rhizosphere predicts the success of root colonization.mSystems · 2025Article
- ATF6 activation alters colonic lipid metabolism causing tumour-associated microbial adaptation.Nature metabolism · 2025Article
- Residue-Specific Incorporation of Noncanonical Amino Acids in Auxotrophic Hosts:Chemical reviews · 2025Review
- Clear as mud redefined: Tunable transparent mineral scaffolds for visualizing microbial processes below ground.PNAS nexus · 2025Article
65 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
Here we describe the application of a new click chemistry method for fluorescent tracking of protein synthesis in individual microorganisms within environmental samples. This technique, termed bioorthogonal non-canonical amino acid tagging (BONCAT), is based on the in vivo incorporation of the non-canonical amino acid L-azidohomoalanine (AHA), a surrogate for l-methionine, followed by fluorescent labelling of AHA-containing cellular proteins by azide-alkyne click chemistry. BONCAT was evaluated with a range of phylogenetically and physiologically diverse archaeal and bacterial pure cultures and enrichments, and used to visualize translationally active cells within complex environmental samples including an oral biofilm, freshwater and anoxic sediment. We also developed combined assays that couple BONCAT with ribosomal RNA (rRNA)-targeted fluorescence in situ hybridization (FISH), enabling a direct link between taxonomic identity and translational activity. Using a methanotrophic enrichment culture incubated under different conditions, we demonstrate the potential of BONCAT-FISH to study microbial physiology in situ. A direct comparison of anabolic activity using BONCAT and stable isotope labelling by nano-scale secondary ion mass spectrometry ((15)NH(3) assimilation) for individual cells within a sediment-sourced enrichment culture showed concordance between AHA-positive cells and (15)N enrichment. BONCAT-FISH offers a fast, inexpensive and straightforward fluorescence microscopy method for studying the in situ activity of environmental microbes on a single-cell level.
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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.