ArticleJournal of biomolecular NMR2015
Affordable uniform isotope labeling with (2)H, (13)C and (15)N in insect cells.
Article in Journal of biomolecular NMR, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- Extended local deuteration of leucine enhances methyl NMR of large proteins expressed in eukaryotic cells.Chemical communications (Cambridge, England) · 2026Article
- Methyl-specific NMR of therapeutic antibodies: cost-effective isotopic labeling strategies in CHO cells for high-resolution structural characterization.Journal of biomolecular NMR · 2026Article
- Human cells for human proteins: Isotope labeling in mammalian cells in suspension for functional NMR studies.Protein science : a publication of the Protein Society · 2026Article
- Cupriavidus necator, an alternative source for isotopic enrichment of proteins expressed in insect cells for NMR investigations.Journal of biomolecular NMR · 2026Article
- Murine Norovirus: Additional Protocols for Basic and Antiviral Studies.Current protocols · 2023Article
- Stabilization of pre-existing neurotensin receptor conformational states by β-arrestin-1 and the biased allosteric modulator ML314.Nature communications · 2023Article
- Specific isotopic labelling and reverse labelling for protein NMR spectroscopy: using metabolic precursors in sample preparation.Biochemical Society transactions · 2022Review
- Local Deuteration Enables NMR Observation of Methyl Groups in Proteins from Eukaryotic and Cell-Free Expression Systems.Angewandte Chemie (International ed. in English) · 2021Article
- GPCR Activation States Induced by Nanobodies and Mini-G Proteins Compared by NMR Spectroscopy.Molecules (Basel, Switzerland) · 2020Article
- Enabling NMR Studies of High Molecular Weight Systems Without the Need for Deuteration: The XL-ALSOFAST Experiment with Delayed Decoupling.Angewandte Chemie (International ed. in English) · 2020Article
- Improved strategy for isoleucineJournal of biomolecular NMR · 2019Article
- Deuteration and selective labeling of alanine methyl groups of βJournal of biomolecular NMR · 2018Article
- Production of isotope-labeled proteins in insect cells for NMR.Journal of biomolecular NMR · 2018Review
- Funaria hygrometrica Hedw. elevated tolerance to DPlanta · 2018Article
- An improved protocol for amino acid type-selective isotope labeling in insect cells.Journal of biomolecular NMR · 2017Article
- Current strategies for protein production and purification enabling membrane protein structural biology.Biochemistry and cell biology = Biochimie et biologie cellulaire · 2016Review
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80% can be achieved for (15)N and (13)C with yields comparable to expression in full media. For (2)H,(15)N and (2)H,(13)C,(15)N labeling, incorporation is only slightly lower with 75 and 73%, respectively, and yields are typically twofold reduced. The media were optimized for isotope incorporation, reproducibility, simplicity and cost. High isotope incorporation levels for all labeling patterns are achieved by using labeled algal amino acid extracts and exploiting well-known biochemical pathways. The final formulation consists of just five commercially available components, at costs 12-fold lower than labeling media from vendors. The approach was applied to several cytosolic and secreted target proteins.
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