TY  - JOUR
AU  - Dubey, Abhinav
AU  - Stoyanov, Nikolay
AU  - Viennet, Thibault
AU  - Chhabra, Sandeep
AU  - Elter, Shantha
AU  - Borggräfe, Jan
AU  - Viegas, Aldino
AU  - Nowak, Radosław P.
AU  - Burdzhiev, Nikola
AU  - Petrov, Ognyan
AU  - Fischer, Eric S.
AU  - Etzkorn, Manuel
AU  - Gelev, Vladimir
AU  - Arthanari, Haribabu
TI  - Local Deuteration Enables NMR Observation of Methyl Groups in Proteins from Eukaryotic and Cell‐Free Expression Systems
JO  - Angewandte Chemie / International edition
VL  - 60
IS  - 25
SN  - 0570-0833
CY  - Weinheim
PB  - Wiley-VCH
M1  - FZJ-2021-05879
SP  - 13783 - 13787
PY  - 2021
AB  - Therapeutically relevant proteins such as GPCRs, antibodies and kinases face clear limitations in NMR studies due to the challenges in site-specific isotope labeling and deuteration in eukaryotic expression systems. Here we describe an efficient and simple method to observe the methyl groups of leucine residues in proteins expressed in bacterial, eukaryotic or cell-free expression systems without modification of the expression protocol. The method relies on simple stereo-selective 13C-labeling and deuteration of leucine that alleviates the need for additional deuteration of the protein. The spectroscopic benefits of “local” deuteration are examined in detail through Forbidden Coherence Transfer (FCT) experiments and simulations. The utility of this labeling method is demonstrated in the cell-free synthesis of bacteriorhodopsin and in the insect-cell expression of the RRM2 domain of human RBM39.
LB  - PUB:(DE-HGF)16
C6  - pmid:33768661
UR  - <Go to ISI:>//WOS:000647987600001
DO  - DOI:10.1002/anie.202016070
UR  - https://juser.fz-juelich.de/record/904309
ER  -