THE USE OF SPIN LABELS IN STUDYING THE MECHANISM OF FUNCTION OF THE TRANSPORT ENZYME NA,K-ATPASE
The mechanism of Na,K-ATPase function was studied using nitroxyl radicals and a paramagnetic ion incorporated into various
regions of the enzyme. The nitroxyl radical PHMB-R and a spin benzocarboline (BC) probe were used as spin labels, while
paramagnetic Mn2+ ions were localized in the active site of Na,K-ATPase.The introduction of spin labels into the protein molecule
outside the active site allows for the study of long-range effects and the determination of their location in the membrane. In this
study, the distance between Mn2+ ions in the enzyme's substrate site and the nitroxyl PCMB-R radical, as well as between Mn2+
ions and the spin BC probe in spin-labeled Na,K-ATPase was determined. EPR spectra were recorded in frozen spin-labeled Na,K-
ATPase samples using the continuous saturation method at 77 K.
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