Deuteration of proteins boosted by cell lysates: high-resolution amide and Hα MAS NMR without re-protonation bottleneck
Federico Napoli,
Jia-Ying Guan,
Charles-Adrien Arnaud
et al.
Abstract:Amide-proton detected magic-angle spinning NMR of deuterated proteins has become a main technique in NMR-based structural biology. In standard deuteration protocols that rely on D2O-based culture media, non-exchangeable amide sites remain deuterated, making these sites unobservable. Here we demonstrate that proteins produced with H2O-based culture medium doped with deuterated cell lysate allow to overcome this “reprotonation bottleneck”, while retaining a high level of deuteration (ca. 80 %) and narrow line wi… Show more
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