Effect of Cd(2+) and Hg(2+) on the activity of Na(+)/K(+)-ATPase and Mg(2+)-ATPase adsorbed on polystyrene microtiter plates.
Anal Biochem
; 300(2): 113-20, 2002 Jan 15.
Article
en En
| MEDLINE
| ID: mdl-11779101
ABSTRACT
In the present study a polystyrene microtiter plate was tested as a support material for synaptic plasma membrane (SPM) immobilization by adsorption. The adsorption was carried out by an 18-h incubation at +4 degrees C of SPM with a polystyrene matrix, at pH 7.4. Evaluation of the efficiency of the applied immobilization method revealed that 10% protein fraction of initially applied SPM was bound to the support and that two SPM enzymes, Na(+)/K(+)-ATPase and Mg(2+)-ATPase, retained 70-80% activity after the adsorption. In addition, adsorption stabilizes Na(+)/K(+)-ATPase and Mg(2+)-ATPase, since the activities are substantial 3 weeks after the adsorption. Parallel kinetic analysis showed that adsorption does not alter significantly the kinetic properties of Na(+)/K(+)-ATPase and Mg(2+)-ATPase and their sensitivity to and mechanism of Cd(2+)- or Hg(2+)-induced inhibition. The only exception is the "high affinity" Mg(2+)-ATPase moiety, whose affinity for ATP and sensitivity toward Cd(2+) were increased by the adsorption. The results show that such system may be used as a practical and comfortable model for the in vitro toxicological investigations.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Poliestirenos
/
Cadmio
/
ATPasa de Ca(2/) y Mg(2/)
/
ATPasa Intercambiadora de Sodio-Potasio
/
Enzimas Inmovilizadas
/
Mercurio
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Anal Biochem
Año:
2002
Tipo del documento:
Article
País de afiliación:
Yugoslavia