Secondary and tertiary structure of nucleotide-binding domain of alphasubunit of Na+/K+-ATPase.
The nucleotide-binding domain of the alpha subunit of mouse brain Na+/K+-ATPase was expressed and isolated from Escherichia coli cells. A model structure was constructed by comparative modeling with and without docked ATP. This was compared with the secondary structure determination from UV circular dichroism and Raman spectroscopy. Thus, we support the quality of the model and the correct folding of the recombinant protein. ATP binding was followed by Raman difference spectroscopy, and its influence on the secondary structure of the N domain seems to not be significant.
Hofbauerová
Katerina
K
Department of Physical and Macromolecular Chemistry, Charles University, Albertov 2030, CZ-128 40 Prague 2, Czech Republic. hofbauer@biomed.cas.cz
Kopecký
Vladimír
V
Ettrich
Rüdiger
R
Ettrichová
Olga
O
Amler
Evzen
E
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry* / Q000378:metabolism; D013059:Spectrum Analysis, Raman
D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase