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Analysis of membrane fractions from Mycoplasma gallisepticum
Authors:Christian le Grimellec  Max Zollinger  Marie-Cecile Giocondi
Affiliation:Department of Biochemistry and Maisonneuve-Rosemont Research Center, University of Montreal, Montreal, Quebec H3C 3J7 Canada
Abstract:Membrane fractions have been isolated from Mycoplasma gallisepticum following a procedure derived from that described by Maniloff, J. and Quinlan, D.C. (J. Bacteriol. (1974) 120, 495–501). A light fraction F1 was obtained which contained structures resembling the bleb-infrableb apparatus characteristic of M. gallisepticum. It was enriched in DNA and had an electrophoretic profile different from that of unfractionated membranes. Cholesterol-to-phospholipid ratios higher than two and elevated values of the ratio of saturated to unsaturated fatty acids were other characteristics of this fraction. The two other fractions isolated (FII and FIV) also differed from intact membranes by their cholesterol and phospholipid content as well as by their saturation ratios. The membrane fluidity of FII and FIV, estimated by fluorescence polarization, was similar to that of unfractionated membranes while a slight but significant difference was recorded for the light fraction. Possible relationships between the lateral heterogeneity of the M. gallisepticum membrane and the obtainment of fractions are discussed.
Keywords:Membrane fractionation  Lipid composition  Membrane fluidity  Fluorescence polarization  (M. gallisepticum)  DPH  1,6-diphenyl-1,3,5-hexatriene  β-buffer  0.15 M NaCl/0.01 M 2-mercaptoethanol/0.05 M Tris-HCl, pH 7.4
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