ArticleEuropean biophysics journal : EBJ2008
Selective deuteration of tryptophan and methionine residues in maltose binding protein: a model system for neutron scattering.
Article in European biophysics journal : EBJ, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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15 citing papers in PubMed, 28 citations in OpenAlex.
- Structural characterization of an intrinsically disordered protein complex using integrated small-angle neutron scattering and computing.Protein science : a publication of the Protein Society · 2023Article
- Article
- Production of perdeuterated fucose from glyco-engineered bacteria.Glycobiology · 2021Article
- Synthesis of 2-D-Tetrahedron · 2019Article
- The Production of Matchout-Deuterated Cholesterol and the Study of Bilayer-Cholesterol Interactions.Scientific reports · 2019Article
- Matchout deuterium labelling of proteins for small-angle neutron scattering studies using prokaryotic and eukaryotic expression systems and high cell-density cultures.European biophysics journal : EBJ · 2017Article
- Investigating Structure and Dynamics of Proteins in Amorphous Phases Using Neutron Scattering.Computational and structural biotechnology journal · 2017Review
- Neutrons describe ectoine effects on water H-bonding and hydration around a soluble protein and a cell membrane.Scientific reports · 2016Article
- Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli.Applied microbiology and biotechnology · 2015Article
- Solution conformations of early intermediates in Mos1 transposition.Nucleic acids research · 2013Article
- Neutron fibre diffraction studies of amyloid using H2O/D2O isotopic replacement.Acta crystallographica. Section F, Structural biology and crystallization communications · 2011Article
- Interfacial assembly of proteins and peptides: recent examples studied by neutron reflection.Journal of the Royal Society, Interface · 2009Review
- Preliminary neutron crystallographic analysis of selectively CH3-protonated deuterated rubredoxin from Pyrococcus furiosus.Acta crystallographica. Section F, Structural biology and crystallization communications · 2008Article
- A preliminary neutron crystallographic study of thaumatin.Acta crystallographica. Section F, Structural biology and crystallization communications · 2008Article
- New sources and instrumentation for neutrons in biology.Chemical physicsArticle
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Authors and funding
6 authors at 3 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe methods that have been developed within the ILL-EMBL Deuteration Laboratory for the production of maltose binding protein (MBP) that has been selectively labelled either with deuterated tryptophan or deuterated methionine (single labelling), or both (double labelling). MBP is used as an important model system for biophysical studies, and selective labelling can be helpful in the analysis of small-angle neutron scattering (SANS) data, neutron reflection (NR) data, and high-resolution neutron diffraction data. The selective labelling was carried out in E. coli high-cell density cultures using auxotrophic mutants in minimal medium containing the required deuterated precursors. Five types of sample were prepared and studied: (1) unmodified hydrogenated MBP (H-MBP), (2) perdeuterated MBP (D-MBP), (3) singly labelled MBP with the tryptophan residues deuterated (D-trp MBP), (4) singly labelled MBP with methionine residues deuterated (D-met MBP) and (5) doubly labelled MBP with both tryptophan and methionine residues deuterated (D-trp/met MBP). Labelled samples were characterised by size exclusion chromatography, gel electrophoresis, light scattering and mass spectroscopy. Preliminary small-angle neutron scattering (SANS) experiments have also been carried out and show measurable differences between the SANS data recorded for the various labelled analogues. More detailed SANS experiments using these labelled MBP analogues are planned; the degree to which such data could enhance structure determination by SANS is discussed.
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