Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation
Abstract The vitamin D metabolite, 24 R ,25‐dihydroxyvitamin D 3 (24 R ,25(OH) 2 D 3 ), was tested for its ability to specifically bind to basal lateral membranes isolated from intestinal epithelium of Atlantic cod (a seawater fish), carp (a freshwater fish), and chicken. Specific saturable binding...
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crwiley:10.1002/jcb.1229 2024-06-02T08:03:06+00:00 Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation Larsson, Dennis Nemere, Ilka Sundell, Kristina 2001 http://dx.doi.org/10.1002/jcb.1229 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fjcb.1229 https://onlinelibrary.wiley.com/doi/pdf/10.1002/jcb.1229 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Journal of Cellular Biochemistry volume 83, issue 2, page 171-186 ISSN 0730-2312 1097-4644 journal-article 2001 crwiley https://doi.org/10.1002/jcb.1229 2024-05-03T11:48:49Z Abstract The vitamin D metabolite, 24 R ,25‐dihydroxyvitamin D 3 (24 R ,25(OH) 2 D 3 ), was tested for its ability to specifically bind to basal lateral membranes isolated from intestinal epithelium of Atlantic cod (a seawater fish), carp (a freshwater fish), and chicken. Specific saturable binding was demonstrated in membranes from all three species. Membranes from Atlantic cod, carp, and chicken revealed K d 's of 7.3 ± 0.9, 12.5 ± 0.9 and 7.8 ± 0.1 nM, and a B max for each species estimated to 57.9 ± 2.9, 195.1 ± 8.4 and 175 ± 0.8 fmol/mg protein, respectively. Scatchard analyses indicated a convex curvature and Hill analyses revealed apparent Hill coefficients of 1.84 ± 0.28, 1.80 ± 0.29, and 1.78 ± 0.27 for Atlantic cod, carp and chicken, suggesting a positive cooperative binding in all three species. Basal lateral membranes from Atlantic cod and carp were used to further characterize the binding moiety. In competition studies, basal lateral membranes from Atlantic cod or carp did not discriminate between 24 R ,25(OH) 2 D 3 and the 24 S ,25(OH) 2 D 3 isomer, whereas, 1,25(OH) 2 D 3 and 25(OH)D 3 , were less effective in competing with [ 3 H]24 R ,25(OH) 2 D 3 for binding to basal lateral membranes in Atlantic cod and carp. In both the Atlantic cod and carp enterocyte basal lateral membranes, the binding activity could be extracted equally well with high salt as with detergent, indicating a peripheral membrane protein rather than an integral membrane binding protein. Finally, isolated Atlantic cod and carp enterocytes were chosen for analyses of signal transduction events mediated by the putative receptor. In both species, 24 R ,25(OH) 2 D 3 but not 24 S ,25(OH) 2 D 3 , suppressed Ca 2+ ‐uptake by enterocytes in a dose‐dependent manner. Enterocytes from Atlantic cod and carp, acclimated to Ca 2+ ‐free media, responded by an intracellular Ca 2+ ‐release within seconds after addition of 24 R ,25(OH) 2 D 3 or 24 S ,25(OH) 2 D 3 . The effects on intracellular Ca 2+ ‐release were dose‐dependent for both ... Article in Journal/Newspaper atlantic cod Wiley Online Library Journal of Cellular Biochemistry 83 2 171 186 |
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English |
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Abstract The vitamin D metabolite, 24 R ,25‐dihydroxyvitamin D 3 (24 R ,25(OH) 2 D 3 ), was tested for its ability to specifically bind to basal lateral membranes isolated from intestinal epithelium of Atlantic cod (a seawater fish), carp (a freshwater fish), and chicken. Specific saturable binding was demonstrated in membranes from all three species. Membranes from Atlantic cod, carp, and chicken revealed K d 's of 7.3 ± 0.9, 12.5 ± 0.9 and 7.8 ± 0.1 nM, and a B max for each species estimated to 57.9 ± 2.9, 195.1 ± 8.4 and 175 ± 0.8 fmol/mg protein, respectively. Scatchard analyses indicated a convex curvature and Hill analyses revealed apparent Hill coefficients of 1.84 ± 0.28, 1.80 ± 0.29, and 1.78 ± 0.27 for Atlantic cod, carp and chicken, suggesting a positive cooperative binding in all three species. Basal lateral membranes from Atlantic cod and carp were used to further characterize the binding moiety. In competition studies, basal lateral membranes from Atlantic cod or carp did not discriminate between 24 R ,25(OH) 2 D 3 and the 24 S ,25(OH) 2 D 3 isomer, whereas, 1,25(OH) 2 D 3 and 25(OH)D 3 , were less effective in competing with [ 3 H]24 R ,25(OH) 2 D 3 for binding to basal lateral membranes in Atlantic cod and carp. In both the Atlantic cod and carp enterocyte basal lateral membranes, the binding activity could be extracted equally well with high salt as with detergent, indicating a peripheral membrane protein rather than an integral membrane binding protein. Finally, isolated Atlantic cod and carp enterocytes were chosen for analyses of signal transduction events mediated by the putative receptor. In both species, 24 R ,25(OH) 2 D 3 but not 24 S ,25(OH) 2 D 3 , suppressed Ca 2+ ‐uptake by enterocytes in a dose‐dependent manner. Enterocytes from Atlantic cod and carp, acclimated to Ca 2+ ‐free media, responded by an intracellular Ca 2+ ‐release within seconds after addition of 24 R ,25(OH) 2 D 3 or 24 S ,25(OH) 2 D 3 . The effects on intracellular Ca 2+ ‐release were dose‐dependent for both ... |
format |
Article in Journal/Newspaper |
author |
Larsson, Dennis Nemere, Ilka Sundell, Kristina |
spellingShingle |
Larsson, Dennis Nemere, Ilka Sundell, Kristina Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation |
author_facet |
Larsson, Dennis Nemere, Ilka Sundell, Kristina |
author_sort |
Larsson, Dennis |
title |
Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation |
title_short |
Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation |
title_full |
Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation |
title_fullStr |
Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation |
title_full_unstemmed |
Putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin D 3 in carp and Atlantic cod enterocytes: Characterization of binding and effects on intracellular calcium regulation |
title_sort |
putative basal lateral membrane receptors for 24,25‐dihydroxyvitamin d 3 in carp and atlantic cod enterocytes: characterization of binding and effects on intracellular calcium regulation |
publisher |
Wiley |
publishDate |
2001 |
url |
http://dx.doi.org/10.1002/jcb.1229 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fjcb.1229 https://onlinelibrary.wiley.com/doi/pdf/10.1002/jcb.1229 |
genre |
atlantic cod |
genre_facet |
atlantic cod |
op_source |
Journal of Cellular Biochemistry volume 83, issue 2, page 171-186 ISSN 0730-2312 1097-4644 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/jcb.1229 |
container_title |
Journal of Cellular Biochemistry |
container_volume |
83 |
container_issue |
2 |
container_start_page |
171 |
op_container_end_page |
186 |
_version_ |
1800747560722235392 |