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Composition of axolemma-enriched fractions isolated from bovine CNS myelinated axons

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Abstract

Axolemma-enriched fractions were isolated from the white matter of bovine corpus callosum via a purified preparation of myelinated axons which were osmotically shocked and fractionated on a discontinuous density gradient. Two membrane fractions of differing density were obtained; both were somewhat enriched over white matter whole homogenate in specific activity of acetylcholinesterase and 5′-nucleotidase and maximal binding capacity for saxitoxin. Both membrane fractions contained appreciable amounts of 2′, 3′-cyclic nucleotide 3′-phospho-hydrolase; the specific activity of antimycin-sensitive NAPH-cytochromec reductase and cytochromec oxidase indicated low levels of contamination by microsomal and mitochondrial membrane. The myelin which is concomitantly isolated with the axolemma-enriched fractions has a lipid and protein composition comparable to that of myelin isolated by other procedures. Both axolemma-enriched fractions contain about one half of their dry weight as lipid comprised of approximately 25% cholesterol, 25% galactolipid (cerebrosides and sulfatides in a molar ratio of about 4:1) and 50% phospholipid, mostly choline phosphatides and ethanolamine phospholes in an equimolar ratio. The axolemma fractions are also enriched in ganglioside content relative to the myelin fraction. The polypeptides of the axolemma-enriched fractions range from 20,000 to over 200,000 in molecular weight; the predominant proteins are in the range from 50,000 to 69,000. The most dense axolemma-enriched fraction is over fourfold enriched in glyco-protein content compared with myelin, with at least 10 different molecular-weight classes of glycoproteins as identified by Schiff stain of polyacrylamide gel protein profiles. The differences and similarities in the molecular composition of axolemma-enriched preparations which have been characterized to date are discussed.

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  1. Department of Biochemistry, Medical College of Virginia, Richmond, Virginia

    George H. DeVries, William Payne & Richard G. Saul

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  2. William Payne
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  3. Richard G. Saul
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DeVries, G.H., Payne, W. & Saul, R.G. Composition of axolemma-enriched fractions isolated from bovine CNS myelinated axons. Neurochem Res 6, 521–537 (1981). https://doi.org/10.1007/BF00964391

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