Journal of the Serbian Chemical Society 2002 Volume 67, Issue 12, Pages: 809-817
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Immobilization of Na, K-ATPase isolated from rat brain synaptic plasma membranes
Momić Tatjana, Vujčić Zoran M., Vasić Vesna M., Horvat Anica I.
Rat brain Na,K-ATPase partially purified by SDS from synaptic plasma membranes (SPM) was immobilized by adsorption on nitrocellulose (NC) polyvinylidene fluoride (PVDF) and glass fiber (GF) membranes. Partial SDS solubilization increased the enzyme activity by 40%. With regard to the preservation of the enzyme activity, nitrocellulose was shown to be the optimal support for the immobilization. The enzyme showed the highest percentage activity (14%) after 30 min of SPM adsorption, at 20ºC under the vaccum, with 25 μg of proteins per NC disc filter. In addition adsorption on NC stabilizes the Na,K-ATPase, since the activity was substantial 72 h after adsorption at 20ºC. After adsorption, the sensitivity of the enzyme to HgCl2 and CdCl2 inhibition was higher. The results show that immobilized Na,K-ATPase SPMcan be used as a practical model for the detection of metal ions in different samples.
Keywords: Na, K-ATPase, synaptic plasma membranes, immobilization, inhibition, cadmium, mercury.