13CHD2 methyl group probes of millisecond time scale exchange in proteins by 1H relaxation dispersion: an application to proteasome gating residue dynamics.

Journal article

A pulse scheme is presented for quantifying millisecond time scale chemical exchange processes in proteins by measuring (1)H CPMG relaxation dispersion profiles of (13)CHD(2) methyl groups. The use of (13)CHD(2) isotopomers for (1)H methyl dispersion experiments eliminates problems with interconversion between differentially relaxing proton transitions that complicate the extraction of accurate exchange parameters when (13)CH(3) probes are used. Good agreement is demonstrated between extracted chemical shift differences from fits of dispersion profiles and the corresponding differences measured independently on a model exchanging system, validating the experiment. The methodology is applied to the gating residues of the T. acidiphilium proteasome that are shown to undergo extensive motion on the millisecond time scale.

Authors: Baldwin, A; Religa, T; Hansen, D; Bouvignies, G; Kay, L

• Publication status: Published
• Journal: Journal of the American Chemical Society
• Volume: 132
• Issue: 32
• Pages: 10992-10995
• Publication date: 2010-08-01
• DOI: 10.1021/ja104578n
• EISSN: 1520-5126
• ISSN: 0002-7863
• Language: English
• Keywords: Methane; Proteasome Endopeptidase Complex; Nuclear Magnetic Resonance, Biomolecular; Thermoplasma; Protein Conformation; Kinetics; Deuterium; Models, Molecular