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Solid‐State NMR Studies of Adenosine 5′‐Triphosphate Freeze‐Trapped in the Nucleotide Site of Na,K‐ATPase
Author(s) -
Middleton David A.,
Hughes Eleri,
Fedosova Natalya U.,
Esmann Mikael
Publication year - 2009
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200900167
Subject(s) - nucleotide , adenosine triphosphate , atpase , atp hydrolysis , nuclear magnetic resonance spectroscopy , substrate (aquarium) , chemistry , enzyme , biochemistry , biophysics , crystallography , stereochemistry , biology , gene , ecology
Freeze frame : The ubiquitous ion pump, Na,K‐ATPase (NKA), was discovered over half a century ago, yet atomic details of the native substrate ATP in the nucleotide site remain unavailable. Here, we use low‐temperature solid‐state NMR spectroscopy to detect uniformly 13 C‐labelled ATP bound to NKA in membrane preparations. Freeze‐trapping the complex prevents ATP hydrolysis and allows the detection of specific nucleotide–enzyme contacts.