ATP-binding is stabilized by a stacking interaction within the binding site of Na(+)/K(+)-ATPase

0142459 - FGU-C 20030064 RIV US eng J - Journal Article
Hofbauerová, Kateřina - Kopecký ml., Vladimír - Ettrich, Rüdiger - Kubala, Martin - Teisinger, Jan - Amler, Evžen
ATP-binding is stabilized by a stacking interaction within the binding site of Na(+)/K(+)-ATPase.
Biochemical and Biophysical Research Communications. Roč. 306, č. 2 (2003), s. 416-420. ISSN 0006-291X. E-ISSN 1090-2104
R&D Projects: GA ČR GA204/01/0254; GA ČR GA204/01/1001; GA ČR GA309/02/1479
Institutional research plan: CEZ:AV0Z5011922; CEZ:MSM 113100001; CEZ:MSM 113200001; CEZ:MSM 123100001; CEZ:AV0Z3042911
Keywords : ATP-binding site * Na(+)/K(+)-ATPase * moledular modeling
Subject RIV: BO - Biophysics
Impact factor: 2.836, year: 2003

Site-directed mutagenesis was applied to modify phenylalanines (Phe(475)Trp, Phe(548)Tyr and both) to generate mutants on the basis of molecular modeling of the ATP-binding domain of Na(+)/K(+)-ATPase, in order to characterize the forces that stabilize ATP in its binding pocket. Each of the mutants was examined by Raman difference spectroscopy, i.e. as a difference between the spectrum of the domain with and without bound ATP. It was shown that Phe(475) plays a key role in stabilizing ATP-binding by a stacking interaction. Phe(548) co-stabilizes ATP on the opposite site of the binding pocket and its type of interaction with ATP-binding differs from that of Phe(475).
Permanent Link: http://hdl.handle.net/11104/0040162