Effect of Cd2+ and Hg2+ on the activity of Na+/K+-ATPase and Mg2+-ATPase adsorbed on polystyrene microtiter plates
Apstrakt
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 +4degreesC 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 Mg2+-ATPase, retained 70-80% activity after the adsorption. In addition, adsorption stabilizes Na+/K+-ATPase and Mg2+-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 Mg2+-ATPase and their sensitivity to and mechanism of Cd2+- or Hg2+- induced inhibition. The only exception is the high affinity Mg2+-ATPase moiety, whose affinity for ATP and sensitivity toward Cd2+ were increased by the adsorp...tion. The results show that such system may be used as a practical and comfortable model for the in vitro toxicological investigations. (C) 2001 Elsevier Science.
Ključne reči:
Na+/K+-ATPase / Mg2+-ATPase / synaptic plasma membranes / Adsorption / heavy metals / kineticsIzvor:
Analytical Biochemistry, 2002, 300, 2, 113-120
DOI: 10.1006/abio.2001.5405
ISSN: 0003-2697
PubMed: 11779101
WoS: 000173371000002
Scopus: 2-s2.0-0037080992
Kolekcije
Institucija/grupa
VinčaTY - JOUR AU - Vasić, Vesna M. AU - Jovanovic, D AU - Horvat, Anica AU - Momić, Tatjana AU - Nizezic, G PY - 2002 UR - https://vinar.vin.bg.ac.rs/handle/123456789/2496 AB - 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 +4degreesC 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 Mg2+-ATPase, retained 70-80% activity after the adsorption. In addition, adsorption stabilizes Na+/K+-ATPase and Mg2+-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 Mg2+-ATPase and their sensitivity to and mechanism of Cd2+- or Hg2+- induced inhibition. The only exception is the high affinity Mg2+-ATPase moiety, whose affinity for ATP and sensitivity toward Cd2+ 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. (C) 2001 Elsevier Science. T2 - Analytical Biochemistry T1 - Effect of Cd2+ and Hg2+ on the activity of Na+/K+-ATPase and Mg2+-ATPase adsorbed on polystyrene microtiter plates VL - 300 IS - 2 SP - 113 EP - 120 DO - 10.1006/abio.2001.5405 ER -
@article{ author = "Vasić, Vesna M. and Jovanovic, D and Horvat, Anica and Momić, Tatjana and Nizezic, G", year = "2002", 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 +4degreesC 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 Mg2+-ATPase, retained 70-80% activity after the adsorption. In addition, adsorption stabilizes Na+/K+-ATPase and Mg2+-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 Mg2+-ATPase and their sensitivity to and mechanism of Cd2+- or Hg2+- induced inhibition. The only exception is the high affinity Mg2+-ATPase moiety, whose affinity for ATP and sensitivity toward Cd2+ 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. (C) 2001 Elsevier Science.", journal = "Analytical Biochemistry", title = "Effect of Cd2+ and Hg2+ on the activity of Na+/K+-ATPase and Mg2+-ATPase adsorbed on polystyrene microtiter plates", volume = "300", number = "2", pages = "113-120", doi = "10.1006/abio.2001.5405" }
Vasić, V. M., Jovanovic, D., Horvat, A., Momić, T.,& Nizezic, G.. (2002). Effect of Cd2+ and Hg2+ on the activity of Na+/K+-ATPase and Mg2+-ATPase adsorbed on polystyrene microtiter plates. in Analytical Biochemistry, 300(2), 113-120. https://doi.org/10.1006/abio.2001.5405
Vasić VM, Jovanovic D, Horvat A, Momić T, Nizezic G. Effect of Cd2+ and Hg2+ on the activity of Na+/K+-ATPase and Mg2+-ATPase adsorbed on polystyrene microtiter plates. in Analytical Biochemistry. 2002;300(2):113-120. doi:10.1006/abio.2001.5405 .
Vasić, Vesna M., Jovanovic, D, Horvat, Anica, Momić, Tatjana, Nizezic, G, "Effect of Cd2+ and Hg2+ on the activity of Na+/K+-ATPase and Mg2+-ATPase adsorbed on polystyrene microtiter plates" in Analytical Biochemistry, 300, no. 2 (2002):113-120, https://doi.org/10.1006/abio.2001.5405 . .