Purification and characterization of chick intestine brush border membrane. Effects of 1alpha(OH) vitamin D3 treatment.
A new technique has been developed for the isolation of membrane vesicles from the vitamin D-deficient and vitamin D-treated chick intestinal brush border membrane. The technique involves removal of nuclei from a low speed pellet by discontinuous sucrose gradient centrifugation. The resulting intact brush borders are then homogenized in 0.5 M Tris and the membrane fragments purified on a glycerol gradient. This preparation represents a 20-fold purification of the brush border marker sucrase. After 1alpha-hydroxyvitamin D3 treatment there is a significant increase in membrane phospholipid phosphorous, an alteration in the fatty acid composition of the phosphatidylcholine fraction of membrane phospholipid, and a decrease in sucrase specific activity.
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects*; D002499:Centrifugation, Density Gradient; D002645:Chickens; D005227:Fatty Acids / Q000032:analysis; D006651:Histocytochemistry; D006887:Hydroxycholecalciferols / Q000494:pharmacology*; D007413:Intestinal Mucosa / Q000648:ultrastructure; D008871:Microvilli / Q000187:drug effects*; D010743:Phospholipids / Q000032:analysis; D013395:Sucrose
D005227:Fatty Acids; D006887:Hydroxycholecalciferols; D010743:Phospholipids; D013395:Sucrose