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Asami, S.* ; Reif, B.

Accessing methyl groups in proteins via 1H-detected MAS solid-state NMR spectroscopy employing random protonation.

Sci. Rep. 9:15903 (2019)
Publ. Version/Full Text Research data DOI PMC
Open Access Gold
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We recently introduced RAP (reduced adjoining protonation) labelling as an easy to implement and cost-effective strategy to yield selectively methyl protonated protein samples. We show here that even though the amount of H2O employed in the bacterial growth medium is rather low, the intensities obtained in MAS solid-state NMR H-1,C-13 correlation spectra are comparable to spectra obtained for samples in which alpha-ketoisovalerate was employed as precursor. In addition to correlations for Leu and Val residues, RAP labelled samples yield also resonances for all methyl containing side chains. The labelling scheme has been employed to quantify order parameters, together with the respective asymmetry parameters. We obtain a very good correlation between the order parameters measured using a GlcRAP (glucose carbon source) and a alpha-ketoisovalerate labelled sample. The labelling scheme holds the potential to be very useful for the collection of long-range distance restraints among side chain atoms. Experiments are demonstrated using RAP and alpha-ketoisovalerate labelled samples of the alpha-spectrin SH3 domain, and are applied to fibrils formed from the Alzheimer's disease A beta(1-40) peptide.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Atomic-resolution Structure; Side-chain Protons; Perdeuterated Proteins; Deuterated Proteins; Molecular-structure; Backbone Dynamics; Labeling Strategy; Dipolar Couplings; Order Parameters; Aliphatic Sites
ISSN (print) / ISBN 2045-2322
e-ISSN 2045-2322
Journal Scientific Reports
Quellenangaben Volume: 9, Issue: 1, Pages: , Article Number: 15903 Supplement: ,
Publisher Nature Publishing Group
Publishing Place London
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Structural Biology (STB)