Opposing Effects of Na+ and K+ on the Thermal Stability of Na+,K+-ATPase. 116(10): 3421–3429

SB. KAUFMAN; FL. GONZÁLEZ FLECHA; GONZÁLEZ-LEBRERO, RODOLFO MARTIN

JOURNAL OF PHYSICAL CHEMISTRY B - (Print)

AMER CHEMICAL SOC

Lugar: Washington; Año: 2012 vol. 116 p. 3421 - 3429

1520-6106

<!-- @page { margin: 2cm } P { margin-bottom: 0.21cm } --> Folding and structural stability are key factors for the proper biological function of proteins. Na+,K+ ATPase is an integral membrane protein involved in the active transport of Na+ and K+ across the plasma membrane. In this work we characterized the effects of K+ and Na+ on the thermal inactivation of Na+,K+ ATPase, evaluating both catalytic and transport capacities of the pump. Both activities of the enzyme decrease with the pre-incubation time as a first-order kinetics. The thermal inactivation of Na+,K+ ATPase is simultaneous with a conformational change detected by tryptophan and ANS fluorescence. The kinetic coefficient of thermal inactivation was affected by the presence of Na+, K+ (or Rb+) and the temperature of the pre-incuabtion media. Our results show that K+ or Rb+ stabilize the enzyme, while Na+ decreases the stability of Na+,K+ ATPase. Both effects are exerted by the specific binding of these cations to the pump. Also, we provided strong evidence that Rb+ (or K+) stabilization effect is due to the occlusion of these cations into the enzyme. Here, we proposed a minimal kinetic model that explains the behavior observed in the experimental results and, allows a better understanding of the results presented by other researchers. The thermal inactivation process was also analyzed according to Kramer’s theory.