Purification and characterization of (Na+ + K+)-ATPase from toad kidney

Détails

ID Serval
serval:BIB_23E29567132B
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Purification and characterization of (Na+ + K+)-ATPase from toad kidney
Périodique
Biochimica et Biophysica Acta (BBA) - Enzymology
Auteur(s)
Geering Kathi, Rossier Bernard C.
ISSN
0005-2744
Statut éditorial
Publié
Date de publication
01/1979
Peer-reviewed
Oui
Volume
566
Numéro
1
Pages
157-170
Langue
anglais
Notes
Publication types: Journal Article - Publication Status: ppublish
Résumé
This report describes the partial purification and the characteristics of (Na+ + K+)-ATPase (ATP phosphohydrolase, EC 3.6.1.3) from an amphibian source. Toad kidney microsomes were solubilized with sodium deoxycholate and further purified by sodium dodecyl sulphate treatment and sucrose gradient centrifugation, according to the methods described by Lane et al. [(1973) J. Biol. Chem. 248, 7197--7200], Jørgensen [(1974) Biochim. Biophys. Acta 356, 36--52] and Hayashi et al. [(1977) Biochim. Biophys. Acta 482, 185--196]. (Na+ + K+)-ATPase preparations with specific activities up to 1000 mumol Pi/mg protein per h were obtained. Mg2+-ATPase only accounted for about 2% of the total ATPase activity. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed three major protein bands with molecular weights of 116 000, 62 000 and 26 000. The 116 000 dalton protein was phosphorylated by [gamma-32P]ATP in the presence of sodium but not in the presence of potassium. The 62 000 dalton component stained for glycoproteins. The Km for ATP was 0.40 mM, for Na+ 12.29 mM and for K+ 1.14 mM. The Ki for ouabain was 35 micron. Temperature activation curves showed two activity peaks at 37 degrees C and at 50 degrees C. The break in the Arrhenius plot of activity versus temperature appeared at 15 degrees C.
Mots-clé
Animals, Anura, Female, Kidney, Kinetics, Male, Microsomes, Molecular Weight, Ouabain, Sodium-Potassium-Exchanging ATPase, Species Specificity
Pubmed
Web of science
Création de la notice
24/01/2008 14:00
Dernière modification de la notice
03/03/2018 14:56
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