Previous studies clearly indicated that membrane enzymes - ATPases i.e. Na, K-ATPase and Mg-ATPase, responded to presence of various organic and inorganic pollutants. In this work effect of mercury and cadmium on these enzymes activities was investigated in synaptic plasma membranes partially immobilized on microliter plate. Comparing those activities with the control enzyme activities obtained with native and partially immobilized mambranes it was concluded that: a. both metals exerted a concentration-dependent inhibition of investigated enzymes, b. for partially immobilized membranes estimated half maximum inhibition (IC50) values for Na, K-ATPase were IC50 (Hg) = 0.9 micromol/cubic cm, IC50 (cd) = 35 micromol/cubic cm and for Mg - ATPase IC50 (Hg) = 3.5 micromol/cubic cm, IC50 (Cd) = 36 micromol/cubic cm; for native membranes IC50 for Na,K-ATPase were IC50 (Hg) =3.3 micromol/cubic cm, IC50 (Cd) = 2 micromol/cubic cm and for Mg ATPase IC50 (Hg) = 2.3 micromol/cubic cm, IC50 (Cd) = 0.2 mmol/cubic cm. Obtained results indicate avaibility of microtitar plates for partially immobilization of membranes with aim to form a new biosensor for heavy metals detection.

Possibilities of application of transmembrane enzymes as a biological component of a biosensor for water quality control and detection of toxical substances were performed. Synaptic plasma membranes (SPMs) were adsorbed on nitrocellulose filters. The adsorption of SPMs was followed by determination of the transmembrane enzyme Na,K-ATPase. The optimal conditions for SPM adsorption on nitrocellulose filters were determined: 25 microgram per nitrocellulose filter disc during 1 hour of incubation, on - 20 deg C. The ATPase activity of adsorbed SPM showed, that almost 30% of enzymic activity was detected on nitrocellulose filters in mentioned conditions. This results showed that adsorption of SPM on solid support enhancing enzymatic stability and enable its industrial and analytical application.

Spectrophotometric method for determination of inorganic phosphate liberated in hydrolysis of ATP catalyzed by ATPase was modified in order to obtain faster procedure, which could also be used in none laboratory conditions. The modification has some advantages compared to the most used Pennial method: a) the reagents are stable for several months; b) the 45 min procedure of phosphomolibdate extraction by isobutanole-benzene is ommited, and the method is not dangerous for the analyst; c) color develops after 20 min. The method was tested on the determination of inorganic phosphate in the presence of cadmium nitrate as inhibitor of ATPase activity. The results were compared to the results obtained by Pennial method. The results obtained have shown some good agreements.

In our earlier work, we have shown that enzymes from rat brain synaptosomal membranes, Na(+)/K(+)-ATPase and Mg(2+)-ATPase, are promising biological components of a biosensor for lead detection. In this work, we represent our results of investigation with the same enzymes as biological components for the biosensor in presence of Hg(2+) ions in water. It was established that IC50 for Na(+)/K(+)-ATPase and Mg(2+)-ATPase is 6.9 and 5.5 x 10E-6 M, and the percentages of inhibition are 96% and 77% respectively. We concluded that these enzymes could be the base for developing biosensors for the presence of Hg(2+) ion in water. Since these enzymes maintain a stable activity for a longer period of time, they could be appropriate as components of biosensors for monitoring water quality.

Activities of rat brain synaptic plasma membrane (SPM)Na(+)/K(+)-ATPase and Mg(2+)-ATPase were investigated by in vitro individual and simultaneous exposure to ions of the first transition series metals (Cu(2+), Zn(2+), Fe(2+)). All investigated metals produced a larger maximum inhibition of Na(+)/K(+)-ATPase than Mg(2+)-ATPase activity. Metal concentrations causing 50% inhibition of Na(+)/K(+)-ATPase activities were: Cu(2+) 7.1 microM Zn(2+) 19 microM Fe(2+) 25 microM. Simulataneous exposure to metal combinations: Cu(2+)/Zn(2+), Cu(2+)/Fe(2+) and Zn(2+)/Fe(2+) inhibited both the Na(+)/K(+)-ATPase and Mg(2+)-ATPase activity synergistically, i.e., more than the sum of the metal induced inhibitions assayed separately. It is also established that the plot of logarithm IC50 values of Na(+)/K(+)-ATPase activity vs. the ionic radius of (Cu(2+), Zn(2+), Fe(2+)) is a straight line that enables to predict the value IC50 for other metals of the first transition series. The results obtained show that Na(+)/K(+)-ATPase is promising as a biological component of biosensor.