Regulation of cellular retinol binding protein type II by 1,25-dihydroxyvitamin D3.
Previously we purified and sequenced an 18-kDa chick duodenal protein that was modulated by 1,25-dihydroxyvitamin D3. The N-terminus of this protein has striking sequence homology to cellular retinol binding protein type II (CRBP II). Furthermore, this purified chick protein binds retinol. Antibodies have now been generated to the chick protein and used for immunoblot analysis to demonstrate that the chick protein has molecular weight, tissue distribution, and subcellular localization similar to rat CRBP II. These antibodies also cross-reacted with rat CRBP II. Antibodies to rat CRBP II cross-react with the chick protein. Northern analysis using a cDNA probe for rat CRBP II showed a single 860 base pair mRNA in both chick and rat intestinal RNA preparations. These results demonstrate that the 1,25-dihydroxyvitamin D3 modulated protein in chick embryonic organ culture is chick CRBP II. Pulse-chase experiments in chick embryonic duodenal organ culture strongly suggest that 1,25-dihydroxyvitamin D3 markedly decreases the synthesis of CRBP II, while not changing the degradation rate. The concentration of 1,25-dihydroxyvitamin D3 required for the decrease in CRBP II synthesis is approximately that required to stimulate calcium uptake into embryonic chick duodenal organ cultures.
Department of Biochemistry, University of Wisconsin--Madison, College of Agricultural and Life Sciences 53706.
J A
JA
Finlay
M
M
Strom
D E
DE
Ong
H F
HF
DeLuca
D000818:Animals; D002117:Calcitriol / Q000494:pharmacology*; D002642:Chick Embryo; D002645:Chickens; D003600:Cytosol / Q000378:metabolism; D004386:Duodenum / Q000378:metabolism; D066298:In Vitro Techniques; D012313:RNA / Q000378:metabolism; D051381:Rats; D012177:Retinol-Binding Proteins / Q000378:metabolism*; D054840:Retinol-Binding Proteins, Cellular
D012177:Retinol-Binding Proteins; D054840:Retinol-Binding Proteins, Cellular; D012313:RNA; D002117:Calcitriol