After extraction of 5'-NT (nucleotidase) from bull seminal plasma vesicles, protein was purified and reconstitued into hen egg yolk lecithin liposomes. Enzyme incorporated into liposomes in a dimeric form with its two subunits linked by disulfie bridges. The incorporation of 5'-NT resulted in the immobilization of the spin probes was demonstrated by electron spin resonance experiment, which performed with liposomes containing electron spin labels localized at the hydrophobic lipid head group (5-Doxyl stearic acid) or in hydrophobic lipid hydrocarbon chain (16-Doxyl stearic acid). Further more the spectral parameters obtained before and after treatment of 5'-NT-containing liposomes with phos-phatidyl inositol phospho lipase C indicated that the liposome membrane bilayer did not contain protein segment.
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