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The frequency-domain method reveals the dimeric structure of Na,K-ATPase
Authors:Ev?en Amler  Roberto Staffolani  Arno?t Kotyk
Institution:(1) Institute of Physiology, Czech Academy of Sciences, Vídencaronská 1083, 142 20 Prague 4, Czech Republic;(2) Institute of Biochemistry, University of Ancona, 601 00 Ancona, Italy
Abstract:Lucifer yellow and lissamine rhodamine sulfonyl hydrazine were used as the donor and the receptor, respectively, for Förster energy transfer measurements to determine the location of the beta subunit in the native Na,K-ATPase from pig kidney. It was found that (1) the beta subunits are located in one functional complex, i.e., the dimer (agrbeta)2 appears to be the functional complex of Na,K-ATPase, and (2) the beta subunits in the functional enzyme complex in the membrane are not located next to each other but are rather well separated. The distance between fluorophores covalently attached to the beta subunits was found to be 5.3 nm.
Keywords:Na  K-ATPase  frequency domain  Lucifer yellow  lissamine rhodamine sulfonylhydrazine  dimeric structure
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