The comparative study of peroxidase activity and substrate binding properties of cytochrome P450 2B4 incorporated into phospholipid vesicles with the use of two different methods

The comparative study of peroxidase activities and substrate binding properties of cytochrome p450 2B4 in reconstituted vesicles prepared with the use of two different techniques, and microsomal cytochrome P450 was carried out. The data obtained show that the two types of cytochrome P450 2B4-containing proteoliposomes do not differ substantially from each other with respect to H2O2- or cumene hydroperoxide-dependent substrate hydroxylation activities as well as substrate binding properties of hemoprotein reconstituted. However, some parameters measured with proteoliposomal cytochrome P450 are markedly different from those measured with microsomal hemoprotein, suggesting the existence of conformational differences between the molecules of these two cytochromes or the differences in the depth of their immersion into lipid bilayer.     

Isolation of a relatively pure outer membrane fraction of Moraxella nonliquefaciens and a comparison of its characteristics with the cytoplasmic membrane-containing material

Cell envelope fractions of Moraxella nonliquefaciens were isolated by a slight modification of Osborn's method. Two main membrane fractions were characterized chemically and morphologically. The density of the fraction containing cytoplasmic membrane material was 1.17 to 1.18 g cm-3 compared with 1.24 to 1.27 g cm-3 for the outer membrane fraction. Lipopolysaccharide was detected almost exclusively in the outer membrane fraction and sodium dodecyl sulphate polyacrylamide gel electrophoresis of this fraction revealed one dominant protein band with an apparent molecular weight of 45 000. Cross-contamination of the fractions was estimated to be about 10%, as calculated on the basis of the lipopolysaccharide fatty acid 3-hydroxydodecanoic acid and on the relative activities of D-lactate dehydrogenase and succinate dehydrogenase.