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. 1985 Dec;76(6):2312–2316. doi: 10.1172/JCI112241

1,25-Dihydroxyvitamin D increases calmodulin binding to specific proteins in the chick duodenal brush border membrane.

D D Bikle, S Munson
PMCID: PMC424356  PMID: 3841129

Abstract

In previous studies we demonstrated that the biologically active vitamin D metabolite 1,25-dihydroxyvitamin D [1,25(OH)2D] increased the calmodulin (CaM) content of chick duodenal brush border membranes (BBM) without increasing the total cellular CaM content. Therefore, we evaluated the binding of CaM to discrete proteins in the BBM and determined whether 1,25(OH)2D could influence such binding. We observed one major and several minor CaM-binding bands on autoradiograms of sodium dodecyl sulfate polyacrylamide gels incubated with [125I]CaM. The major band had a molecular weight of 102,000-105,000. It bound CaM even in the presence of EGTA, but not in the presence of trifluoperazine or excess nonradioactive CaM. The administration of 1,25(OH)2D increased the apparent binding of CaM to this protein as assessed by densitometry of the autoradiogram. This increase in CaM binding coincided with the increased ability of the same BBM vesicles to accumulate calcium. Cycloheximide in doses that markedly reduced the incorporation of [35S]methionine into BBM proteins did not reduce the ability of 1,25-dihydroxyvitamin D3 to stimulate either calcium uptake by the BBM vesicles or CaM binding to the 102,000-105,000-mol-wt protein. These results suggest that 1,25(OH)2D administration increases the CaM content of duodenal BBM by increasing the ability of a 102,000-105,000-mol-wt protein to bind CaM. This mechanism may underlie the ability of 1,25(OH)2D to stimulate calcium movement across the intestinal BBM.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bikle D. D., Askew E. W., Zolock D. T., Morrissey R. L., Herman R. H. Calcium accumulation by chick intestinal mitochondria. Regulation by vitamin D-3 and 1,25-dihydroxyvitamin D-3. Biochim Biophys Acta. 1980 Jun 6;598(3):561–574. doi: 10.1016/0005-2736(80)90036-x. [DOI] [PubMed] [Google Scholar]
  2. Bikle D. D., Morrissey R. L., Zolock D. T., Rasmussen H. The intestinal response to vitamin D. Rev Physiol Biochem Pharmacol. 1981;89:63–142. doi: 10.1007/BFb0035265. [DOI] [PubMed] [Google Scholar]
  3. Bikle D. D., Morrissey R. L., Zolock D. T. The mechanism of action of vitamin D in the intestine. Am J Clin Nutr. 1979 Nov;32(11):2322–2328. doi: 10.1093/ajcn/32.11.2322. [DOI] [PubMed] [Google Scholar]
  4. Bikle D. D., Munson S., Chafouleas J. Calmodulin may mediate 1,25-dihydroxyvitamin D-stimulated intestinal calcium transport. FEBS Lett. 1984 Aug 20;174(1):30–33. doi: 10.1016/0014-5793(84)81071-6. [DOI] [PubMed] [Google Scholar]
  5. Bikle D. D., Munson S., Zolock D. T. Calcium flux across chick duodenal brush border membrane vesicles: regulation by 1,25-dihydroxyvitamin D. Endocrinology. 1983 Dec;113(6):2072–2080. doi: 10.1210/endo-113-6-2072. [DOI] [PubMed] [Google Scholar]
  6. Bikle D. D., Whitney J., Munson S. The relationship of membrane fluidity to calcium flux in chick intestinal brush border membranes. Endocrinology. 1984 Jan;114(1):260–267. doi: 10.1210/endo-114-1-260. [DOI] [PubMed] [Google Scholar]
  7. Bikle D. D., Zolock D. T., Morrissey R. L., Herman R. H. Independence of 1,25-dihydroxyvitamin D3-mediated calcium transport from de novo RNA and protein synthesis. J Biol Chem. 1978 Jan 25;253(2):484–488. [PubMed] [Google Scholar]
  8. COATES M. E., HOLDSWORTH E. S. Vitamin D3 and absorption of calcium in the chick. Br J Nutr. 1961;15:131–147. doi: 10.1079/bjn19610014. [DOI] [PubMed] [Google Scholar]
  9. Collins J. H., Borysenko C. W. The 110,000-dalton actin- and calmodulin-binding protein from intestinal brush border is a myosin-like ATPase. J Biol Chem. 1984 Nov 25;259(22):14128–14135. [PubMed] [Google Scholar]
  10. Coudrier E., Reggio H., Louvard D. Immunolocalization of the 110,000 molecular weight cytoskeletal protein of intestinal microvilli. J Mol Biol. 1981 Oct 15;152(1):49–66. doi: 10.1016/0022-2836(81)90095-4. [DOI] [PubMed] [Google Scholar]
  11. Feher J. J., Wasserman R. H. Studies on the subcellular localization of the membrane-bound fraction of intestinal calcium-binding protein. Biochim Biophys Acta. 1979 Jul 18;585(4):599–610. doi: 10.1016/0304-4165(79)90192-2. [DOI] [PubMed] [Google Scholar]
  12. Glenney J. R., Jr, Glenney P. Comparison of Ca++-regulated events in the intestinal brush border. J Cell Biol. 1985 Mar;100(3):754–763. doi: 10.1083/jcb.100.3.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Glenney J. R., Jr, Glenney P. The microvillus 110K cytoskeletal protein is an integral membrane protein. Cell. 1984 Jul;37(3):743–751. doi: 10.1016/0092-8674(84)90410-0. [DOI] [PubMed] [Google Scholar]
  14. Glenney J. R., Jr, Osborn M., Weber K. The intracellular localization of the microvillus 110K protein, a component considered to be involved in side-on membrane attachment of F-actin. Exp Cell Res. 1982 Mar;138(1):199–205. doi: 10.1016/0014-4827(82)90106-9. [DOI] [PubMed] [Google Scholar]
  15. Glenney J. R., Jr, Weber K. Calmodulin-binding proteins of the microfilaments present in isolated brush borders and microvilli of intestinal epithelial cells. J Biol Chem. 1980 Nov 25;255(22):10551–10554. [PubMed] [Google Scholar]
  16. Howe C. L., Keller T. C., 3rd, Mooseker M. S., Wasserman R. H. Analysis of cytoskeletal proteins and Ca2+-dependent regulation of structure in intestinal brush borders from rachitic chicks. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1134–1138. doi: 10.1073/pnas.79.4.1134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Howe C. L., Mooseker M. S. Characterization of the 110-kdalton actin-calmodulin-, and membrane-binding protein from microvilli of intestinal epithelial cells. J Cell Biol. 1983 Oct;97(4):974–985. doi: 10.1083/jcb.97.4.974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Matsumoto T., Fontaine O., Rasmussen H. Effect of 1,25-dihydroxyvitamin D3 on phospholipid metabolism in chick duodenal mucosal cell. Relationship to its mechanism of action. J Biol Chem. 1981 Apr 10;256(7):3354–3360. [PubMed] [Google Scholar]
  21. Max E. E., Goodman D. B., Rasmussen H. Purification and characterization of chick intestine brush border membrane. Effects of 1alpha(OH) vitamin D3 treatment. Biochim Biophys Acta. 1978 Aug 4;511(2):224–239. doi: 10.1016/0005-2736(78)90316-4. [DOI] [PubMed] [Google Scholar]
  22. Miller A., 3rd, Bronner F. Calcium uptake in isolated brush-border vesicles from rat small intestine. Biochem J. 1981 May 15;196(2):391–401. doi: 10.1042/bj1960391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Morrissey R. L., Zolock D. T., Bikle D. D., Empson R. N., Jr, Bucci T. J. Intestinal response to 1 alpha, 25-dihydroxycholecalciferol. I. RNA polymerase, alkaline phosphatase, calcium and phosphorus uptake in vitro, and in vivo calcium transport and accumulation. Biochim Biophys Acta. 1978 Jan 3;538(1):23–33. doi: 10.1016/0304-4165(78)90248-9. [DOI] [PubMed] [Google Scholar]
  24. Nelson T. Y., Oberwetter J. M., Chafouleas J. G., Boyd A. E., 3rd Calmodulin-binding proteins in a cloned rat insulinoma cell line. Diabetes. 1983 Dec;32(12):1126–1133. doi: 10.2337/diab.32.12.1126. [DOI] [PubMed] [Google Scholar]
  25. Nemere I., Yoshimoto Y., Norman A. W. Calcium transport in perfused duodena from normal chicks: enhancement within fourteen minutes of exposure to 1,25-dihydroxyvitamin D3. Endocrinology. 1984 Oct;115(4):1476–1483. doi: 10.1210/endo-115-4-1476. [DOI] [PubMed] [Google Scholar]
  26. Norman A. W., Putkey J. A., Nemere I. Intestinal calcium transport: pleiotropic effects mediated by vitamin D. Fed Proc. 1982 Jan;41(1):78–83. [PubMed] [Google Scholar]
  27. O'Doherty P. J. 1,25-Dihydroxyvitamin D3 increases the activity of the intestinal phosphatidylcholine deacylation-reacylation cycle. Lipids. 1979 Jan;14(1):75–77. doi: 10.1007/BF02533571. [DOI] [PubMed] [Google Scholar]
  28. Putkey J. A., Spielvogel A. M., Sauerheber R. D., Dunlap C. S., Norman A. W. Vitamin D-mediated intestinal calcium transport. Effects of essential fatty acid deficiency and spin label studies of enterocyte membrane lipid fluidity. Biochim Biophys Acta. 1982 May 21;688(1):177–190. doi: 10.1016/0005-2736(82)90593-4. [DOI] [PubMed] [Google Scholar]
  29. Rasmussen H., Fontaine O., Max E. E., Goodman D. B. The effect of 1alpha-hydroxyvitamin D3 administration on calcium transport in chick intestine brush border membrane vesicles. J Biol Chem. 1979 Apr 25;254(8):2993–2999. [PubMed] [Google Scholar]
  30. Schachter D., Kowarski S. Isolation of the protein IMCal, a vitamin D-dependent membrane component of the intestinal transport mechanism for calcium. Fed Proc. 1982 Jan;41(1):84–87. [PubMed] [Google Scholar]
  31. Tanaka Y., DeLuca H. F., Omdahl J., Holick M. F. Mechanism of action of 1,25-dihydroxycholecalciferol on intestinal calcium transport. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1286–1288. doi: 10.1073/pnas.68.6.1286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thomasset M., Molla A., Parkes O., Demaille J. G. Intestinal calmodulin and calcium-binding protein differ in their distribution and in the effect of vitamin D steroids on their concentration. FEBS Lett. 1981 May 5;127(1):13–16. doi: 10.1016/0014-5793(81)80329-8. [DOI] [PubMed] [Google Scholar]
  33. Verner K., Bretscher A. Microvillus 110K-calmodulin: effects of nucleotides on isolated cytoskeletons and the interaction of the purified complex with F-actin. J Cell Biol. 1985 May;100(5):1455–1465. doi: 10.1083/jcb.100.5.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wasserman R. H., Brindak M. E., Meyer S. A., Fullmer C. S. Evidence for multiple effects of vitamin D3 on calcium absorption: response of rachitic chicks, with or without partial vitamin D3 repletion, to 1,25-dihydroxyvitamin D3. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7939–7943. doi: 10.1073/pnas.79.24.7939. [DOI] [PMC free article] [PubMed] [Google Scholar]

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