Glycopeptide fractions prepared from purified central and peripheral rat myelin.
Myelin was purified from rat brain and sciatic nerve after in vivo labeling with [3H]fucose and [14C]glucosamine to provide a radioactive marker for glycoproteins. The glycoproteins in the isolated myelin were digested exhaustively with pronase, and glycopeptides were isolated from the digest by gel filtration on Bio-Gel P-10. The glycopeptides from brain myelin separated into large and small molecular weight fractions, whereas the glycopeptides of sciatic nerve myelin eluted as a single symmetrical peak. The large and small glycopeptide fractions from central myelin and the single glycopeptide fraction from peripheral myelin were analyzed for carbohydrate by colorimetric and gas liquid chromatographic techniques. The glycopeptides from brain myelin contained 2.4 micrograms of neutral sugar and 0.59 micrograms of sialic acid per mg total myelin protein, whereas sciatic nerve myelin glycopeptides contained 10 micrograms of neutral sugar and 3.8 micrograms of sialic acid per mg total protein. Similarly, the gas-liquid chromatographic analyses showed that the glycopeptides from peripheral myelin contained 4- to 7-fold more of each individual per mg total myelin protein than those from central myelin. Most of the sialic acid and galactose in the glycopeptides from central myelin were in the large molecular weight fraction, and the small molecular weight glycopeptides contained primarily mannose and N-acetylglucosamine. The considerably higher content of glycoprotein-carbohydrate in peripheral myelin supports the results of gel electrophoretic studies, which indicate that the major protein in peripheral myelin is glycosylated while the glycoproteins in purified central myelin are quantitatively minor components.
D000818:Animals; D001921:Brain / Q000378:metabolism; D001923:Brain Chemistry; D005643:Fucose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006020:Glycopeptides / Q000096:biosynthesis / Q000302:isolation & purification; D006601:Hexoses / Q000032:analysis; D008297:Male; D009186:Myelin Sheath / Q000032:analysis*; D009928:Organ Specificity; D051381:Rats; D012584:Sciatic Nerve / Q000032:analysis* / Q000378:metabolism; D012794:Sialic Acids / Q000032:analysis
D006020:Glycopeptides; D006601:Hexoses; D012794:Sialic Acids; D005643:Fucose; D005947:Glucose