A specific binding moiety for 1,25-dihydroxyvitamin D(3) in basal lateral membranes of carp enterocytes.
Carp (Cyprinus carpio), a freshwater fish that lives in a low-calcium environment, and Atlantic cod (Gadus morhua), a seawater fish that lives in a high-calcium environment, were studied for the presence of a novel membrane binding protein ("receptor") for the vitamin D metabolite, 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. Basal lateral membranes from enterocytes of either species were prepared and analyzed for specific saturable binding. Membranes from carp revealed a dissociation constant of 1.23 nM with a maximal binding capacity of 212 fmol/mg protein. In comparison, membranes from Atlantic cod enterocytes revealed very low and nonsignificant levels of specific binding. The [(3)H]1,25(OH)(2)D(3) binding activity in carp was further characterized for protein dependence, detergent extractability, and competition for binding with the metabolites 25(OH)D(3) and 24R,25(OH)(2)D(3). Finally, introduction of 1,25(OH)(2)D(3) to isolated carp enterocytes enhanced protein kinase C activity within 5 min, whereas intracellular Ca(2+) concentrations were unaffected by a range of 1,25(OH)(2)D(3) concentrations, as judged by fura 2 fluorescence. Thus the binding moiety may be a putative plasma membrane receptor for vitamin D, because it is functionally coupled to at least one signal transduction pathway.
Nemere
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I
Department of Nutrition and Food Sciences and the Biotechnology Center, Utah State University, Logan 84322, USA. nemere@cc.usu.edu
Larsson
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D
Sundell
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K
D000818:Animals; D002117:Calcitriol / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002347:Carps / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D020895:Enterocytes / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes; D016257:Fura-2; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000378:metabolism; D007700:Kinetics; D008297:Male; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism; D014809:Vitamin D-Binding Protein / Q000378:metabolism*
D005456:Fluorescent Dyes; D014809:Vitamin D-Binding Protein; D011493:Protein Kinase C; D002117:Calcitriol; D002118:Calcium; D016257:Fura-2