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Solid-state NMR approaches to investigate large enzymes in complex with substrates and inhibitors.

Biochem. Soc. Trans. 49, 131-144 (2021)
DOI Verlagsversion bestellen
Open Access Green: Postprint online verfügbar 01/2022
Enzyme catalysis is omnipresent in the cell. The mechanisms by which highly evolved protein folds enable rapid and specific chemical transformation of substrates belong to the marvels of structural biology. Targeting of enzymes with inhibitors has immediate application in drug discovery, from chemotherapeutics over antibiotics to antivirals. NMR spectroscopy combines multiple assets for the investigation of enzyme function. The non-invasive technique can probe enzyme structure and dynamics and map interactions with substrates, cofactors and inhibitors at the atomic level. With experiments performed at close to native conditions, catalytic transformations can be monitored in real time, giving access to kinetic parameters. The power of NMR in the solid state, in contrast with solution, lies in the absence of fundamental size limitations, which is crucial for enzymes that are either membrane-embedded or assemble into large soluble complexes exceeding hundreds of kilodaltons in molecular weight. Here we review recent progress in solid-state NMR methodology, which has taken big leaps in the past years due to steady improvements in hardware design, notably magic angle spinning, and connect it to parallel biochemical advances that enable isotope labelling of increasingly complex enzymes. We first discuss general concepts and requirements of the method and then highlight the state-of-the-art in sample preparation, structure determination, dynamics and interaction studies. We focus on examples where solid-state NMR has been instrumental in elucidating enzyme mechanism, alone or in integrative studies.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Enzyme Structure ; Enzyme-substrate Interactions ; Isotope Labelling ; Magic Angle Spinning ; Molecular Docking ; Protein Dynamics; Nuclear-magnetic-resonance; Angle-spinning Nmr; Relaxation-dispersion; Protein Assemblies; Tool Box; Spectroscopy; Resolution; Assignment; Dynamics; Polarization
ISSN (print) / ISBN 0300-5127
e-ISSN 1470-8752
Quellenangaben Band: 49, Heft: 1, Seiten: 131-144 Artikelnummer: , Supplement: ,
Verlag Portland Press
Verlagsort 5th Flr, 90 High Holborn, London Wc1v 6lj, England
Begutachtungsstatus Peer reviewed
Förderungen DFG