Vitamin D. Its effect on the protein composition and core material structure of the chick intestinal brush-border membrane.
Vitamin D3 is known to stimulate the absorption of calcium across the asymmetric intestinal epithelial cells. Efforts to elucidate the mechanism of stimulation of intestinal calcium transport by vitamin D are now focused on evaluating the protein composition and topology of the brush-border membrane and its associated core material. Intestinal brush-border membranes were isolated from vitamin D-replete and vitamin D-deficient chicks. Core material proteins were isolated, by sedimentation, from brush-border membranes which were solubilized with Triton X-100. As determined by polyacrylamide gel electrophoresis, dietary vitamin D3 treatment caused no change in the relative amounts of five major core material proteins with Mr = 101,000, 94,000, 67,000, 42,000 (actin), and 17,000. In contrast, dietary vitamin D3 treatment caused a significant reduction in the levels of two proteins with Mr = 111,000 (sucrase) and 83,000, and an increase in the levels of a protein with Mr = 78,000 (possibly a subunit of alkaline phosphatase). The Mr = 111,000, 83,000, and 78,000 proteins are readily solubilized by Triton X-100 and are located on the extracellular surface of the brush-border membrane, as judged by [125I]diazoiodosulfanilic acid and lactoperoxidase 125I labeling. A significant vitamin D-dependent difference was found with respect to iodination of isolated core material as evidenced by the 125I labeling patterns of the Mr = 42,000 protein (actin). The Mr = 42,000 protein was labeled two to three times more extensively when associated with core material derived from vitamin D-deficient chicks as compared to vitamin D-replete chicks. Increasing the salt concentration (0-125 mM KCl) present during core material isolation from either vitamin D-replete or vitamin D-deficient chicks yields core material actin which is more susceptible to iodination by both [125I]diazoiodosulfanilic acid and lactoperoxidase. This increase in the extent of actin iodination is coupled to a salt-induced decrease in the stability of the core material which is evidenced by a decrease in the percentage of total brush-border membrane actin which is Triton-insoluble. This strongly suggests that the vitamin D-induced decrease in the accessibility of actin to iodination reagents results from a vitamin D-dependent change in the structure of the core material. Collectively, these results implicate a role for dietary vitamin D3 in maintaining a specified composition and topology of both the brush-border membrane proteins as well as its associated cytoskeletal core proteins, which is possibly important for intestinal calcium transport.
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002645:Chickens; D002762:Cholecalciferol / Q000494:pharmacology*; D007413:Intestinal Mucosa / Q000378:metabolism*; D007457:Iodine Radioisotopes; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008871:Microvilli / Q000187:drug effects / Q000378:metabolism*; D008970:Molecular Weight; D011189:Potassium Chloride / Q000494:pharmacology; D014808:Vitamin D Deficiency / Q000378:metabolism
D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D002762:Cholecalciferol; D011189:Potassium Chloride