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Detection of nucleotide binding to Na,K‐ATPase in an aqueous membrane suspension by 13 C cross‐polarization magic‐angle spinning NMR spectroscopy
Author(s) -
Middleton David A.,
Jakobsen Louise Odgaard,
Esmann Mikael
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2006.11.026
Subject(s) - magic angle spinning , chemistry , nuclear magnetic resonance spectroscopy , aqueous solution , spinning , polarization (electrochemistry) , analytical chemistry (journal) , spectroscopy , magic angle , nucleotide , crystallography , nuclear magnetic resonance , stereochemistry , chromatography , physics , biochemistry , polymer chemistry , quantum mechanics , gene
Binding of uniformly 13 C labelled ATP to Na,K‐ATPase was studied by 13 C cross‐polarization magic‐angle spinning (CP‐MAS) NMR. In the presence of 30 mM Na + , and with sample‐ and time‐averaging, NMR spectra obtained at 4 °C exhibited several resonances for the bound nucleotide. Chemical shifts suggested that site‐specific changes in the micro‐environment or conformation of the nucleotide occurred in the high affinity binding site. These experiments permit further studies of nucleotide dynamics, structure and binding under physiologically relevant conditions.