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Time-resolved multidimensional NMR with non-uniform sampling

Artikel i vetenskaplig tidskrift
Författare Maxim Mayzel
Joakim Rosenlöw
Linnéa Isaksson
Vladislav Orekhov
Publicerad i Journal of Biomolecular Nmr
Volym 58
Nummer/häfte 2
Sidor 129-139
ISSN 0925-2738
Publiceringsår 2014
Publicerad vid Svenskt NMR-centrum vid Göteborgs universitet
Sidor 129-139
Språk en
Länkar dx.doi.org/10.1007/s10858-013-9811-...
Ämnesord IDP, Post-translational modification, NUS, PARAFAC, MDD, ITAM, CD79, BEST-TROSY, Real-time, PROTEIN-PHOSPHORYLATION, 3-WAY DECOMPOSITION, DATA-COLLECTION, SPECTROSCOPY, RESOLUTION, SPECTRA, RELAXATION, EVENTS, NOESY, CELL
Ämneskategorier Biofysik, Biokemi

Sammanfattning

Time-resolved experiments demand high resolution both in spectral dimensions and in time of the studied kinetic process. The latter requirement traditionally prohibits applications of the multidimensional experiments, which, although capable of providing invaluable information about structure and dynamics and almost unlimited spectral resolution, require too lengthy data collection. Our work shows that the problem has a solution in using modern methods of NMR data collection and signal processing. A continuous fast pulsing three-dimensional experiment is acquired using non-uniform sampling during full time of the studied reaction. High sensitivity and time-resolution of a few minutes is achieved by simultaneous processing of the full data set with the multi-dimensional decomposition. The method is verified and illustrated in realistic simulations and by measuring deuterium exchange rates of amide protons in ubiquitin. We applied the method for characterizing kinetics of in vitro phosphorylation of two tyrosine residues in an intrinsically disordered cytosolic domain of the B cell receptor protein CD79b. Signals of many residues including tyrosines in both phosphorylated and unmodified forms of CD79b are found in a heavily crowded region of 2D H-1-N-15 correlation spectrum and the significantly enhanced spectral resolution provided by the 3D time-resolved approach was essential for the quantitative site-specific analysis.

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