The isolated H4‐H5 cytoplasmic loop of Na,K‐ATPase overexpressed in Escherichia coli retains its ability to bind ATP

FEBS Letters

Obšil, Tomáš²  Mérola, Fabienne¹  Amler, Evžen²  Lewit-Bentley, Anita¹ 

[1] Laboratoire pour l'Utilisation du Rayonnement Electromagnétique, CNRS-MEN-CEA, Bât. 209D, Université Paris Sud, F-91405 Orsay, France;Institute of Physiology, Czech Academy of Sciences, Vı́deňská 1083, Cz-142 20 Prague 4, Czech Republic

关键词: Sodium;    potassium adenosine triphosphatase;    Adenosine triphosphate binding site;    Protein expression;    Etheno-adenosine triphosphate;    Time-resolved fluorescence;    DTT;    dithiothreitol;    GST;    glutathione S-transferase;    H4-H5 loop;    the cytoplasmic loop between transmembrane segments 4 and 5 of the α-subunit of Na;    K-ATPase;    Na;    K-ATPase;    Na+;    K+-transporting ATPase (EC 3.6.1.37);    PITC;    pyrene 5′-isothiocyanate;    TNP-ATP;    2′;    3′-O-trinitrophenyl-ATP;    ci;    pre-exponential amplitude of the ith component;    τi;    lifetime of ith component;    <τ>;    average lifetime;    χ 2 R;    goodness of fit;   

DOI  :  10.1016/S0014-5793(98)00373-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The H₄-H₅ loop of the α-subunit of mouse brain Na,K-ATPase was expressed and isolated from Escherichia coli cells. Using fluorescence analogues of ATP, this loop was shown to retain its capability to bind ATP. Isolation of a soluble H₄-H₅ loop with the native ATP binding site is a crucial step for detailed studies of the molecular mechanism of ATP binding and utilisation.

【 授权许可】

Unknown