Kinetic characterization of Na,K-ATPase from rabbit outer renal medulla: properties of the (alpha beta)(2) dimer

Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
Hérica de Lima Santos, P Ciancaglini

Abstract

We describe and compare the main kinetic characteristics of the (alpha beta)(2) form of rabbit kidney Na,K-ATPase. The dependence of ATPase activity on ATP concentration revealed high (K(0.5)=4 microM) and low (K(0.5)=1.4 mM) affinity sites for ATP, exhibiting negative cooperativity and a specific activity of approximately 700 U/mg. For p-nitrophenylphosphate (PNPP) as substrate, a single saturation curve was found, with a smaller apparent affinity of the enzyme for this substrate (K(0.5)=0.5 mM) and a lower hydrolysis rate (V(M)=42 U/mg). Stimulation of ATPase activity by K(+) (K(0.5)=0.63 mM), Na(+) (K(0.5)=11 mM) and Mg(2+) (K(0.5)=0.60 mM) all showed V(M)'s of approximately 600 U/mg and negative cooperativity. K(+) (K(0.5)=0.69 mM) and Mg(2+) (K(0.5)=0.57 mM) also stimulated PNPPase activity of the (alpha beta)(2) form. Ouabain (K(0.5)=0.01 microM and K(0.5)=0.1 mM) and orthovanadate (K(0.5)=0.06 microM) completely inhibited the ATPase activity of the (alpha beta)(2) form. The kinetic characteristics obtained constitute reference values for diprotomeric (alpha beta)(2)-units of Na,K-ATPase, thus contributing to a better understanding of the biochemical mechanisms of the enzyme.

References

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Citations

Mar 15, 2005·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Prislaine P MagalhãesP Ciancaglini
Mar 1, 2012·Biophysics Reviews·P CiancagliniR G Stabeli
Jan 18, 2006·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·H L SantosP Ciancaglini

Related Concepts

4-aminophenylphosphate, monosilver salt
Na(+)-K(+)-Exchanging ATPase
Striadyne
Aniline Compounds
Metazoa
Hydrolysis
Kidney Papilla
Magnesium
Organopyrophosphorus Compounds
Ouabain

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