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@ARTICLE{Dubey:904309,
author = {Dubey, Abhinav and Stoyanov, Nikolay and Viennet, Thibault
and Chhabra, Sandeep and Elter, Shantha and Borggräfe, Jan
and Viegas, Aldino and Nowak, Radosław P. and Burdzhiev,
Nikola and Petrov, Ognyan and Fischer, Eric S. and Etzkorn,
Manuel and Gelev, Vladimir and Arthanari, Haribabu},
title = {{L}ocal {D}euteration {E}nables {NMR} {O}bservation of
{M}ethyl {G}roups in {P}roteins from {E}ukaryotic and
{C}ell‐{F}ree {E}xpression {S}ystems},
journal = {Angewandte Chemie / International edition},
volume = {60},
number = {25},
issn = {0570-0833},
address = {Weinheim},
publisher = {Wiley-VCH},
reportid = {FZJ-2021-05879},
pages = {13783 - 13787},
year = {2021},
abstract = {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.},
cin = {IBI-7},
ddc = {540},
cid = {I:(DE-Juel1)IBI-7-20200312},
pnm = {5244 - Information Processing in Neuronal Networks
(POF4-524)},
pid = {G:(DE-HGF)POF4-5244},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:33768661},
UT = {WOS:000647987600001},
doi = {10.1002/anie.202016070},
url = {https://juser.fz-juelich.de/record/904309},
}