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• W1569576670 abstract "Abstract Squid retinal nerves, because of their high ratio of axolemma to Schwann cell membrane, were used to develop a procedure to isolate plasma membranes. NADH oxidoreductase and (Na + K)-ATPase were studied as enzyme of plasma membranes in various fractions separated by differential centrifugation of hypotonic homogenates of retinal axons. The pellet which sedimented at 100,000 g (F-100) was selected as a crude plasmalemma fraction because of their highest specific activity for both enzymes. Chemical analysis of F-100 revealed a protein: cholesterol: phospholipids ratio of 37:9:29 as percent of membrane dry weight. RNA and DNA were 0.72 and 0.07%, respectively. Assays of mitochondrial and endoplasmic reticulum enzymes indicated that rare contamination of these structures was present in F-100. Negatively stained electron micrographs of F-100 revealed vesicles formed by concentric membranes. Further purification of F-100 was achieved by linear gradient centrifugatiou which separated two types of membranes. ATPase and NADH oxidoreductase were found in a first protein peak at 37% sucrose (PM 1 ), but mainly ATPase was found in the second peak at 27% sucrose (PM 2 ). Sections of osmium-fixed membranes studied under the electron microscope showed striking density of plasma membranes, 70–80 A in thickness. Proteins from F-100, PM 1 , and PM 2 were solubilized by various procedures and resolved either by sucrose-gradient centrifugation or acrylamide-gel electrophoresis. PM 1 and PM 2 differed in their number of protein bands, as well as in their uv spectrum and pH titration curve. These data together with the EM and the enzymic profile indicate that two types of plasma membranes chemically and structurally different were purified from the retinal axons of the squid. Their probable cellular origin is discussed." @default.
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• W1569576670 title "The molecular organization of nerve membranes" @default.
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• W1569576670 doi "https://doi.org/10.1016/0003-9861(70)90277-8" @default.
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