Expression of cellular retinoic acid binding protein II (chick-CRABP II) in the chick embryo

Retinoic acid-binding protein, which is considered to mediate the biological function of retinoic acid in epithelial differentiation and in the possible control of tumorigenesis, was reproducibly purified from chick-embryo skin by using DEAE-Sephadex and Sephadex G-100 column chromatography and isoelectric focusing. About 1mg of protein was isolated from 60g of skin. The purified protein-ligand complex was found to be homogeneous by electrophoresis on polyacrylamide gels. The binding protein has mol.wt. 17800 and pI 4.5. The binding of [3H]retinoic acid to the protein was completely inhibited by mercury compounds. The inhibition is reversible on treatment without dithithreitol; about 50% of the retinoic acid-binding capacity of the mercury-compound-treated protein is restored by chromatography on Sephadex G-25. iodoacetamide treatment of the protein irreversibly inhibits about 50% of retinoic acid binding.

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Two novel single nucleotide polymorphisms in the promoter of the Cellular Retinoic Acid Binding Protein II gene (CRABP-II)

Molecular and Cellular Probes ◽

10.1016/s0890-8508(02)00110-x ◽

2003 ◽

Vol 17 (1) ◽

pp. 21-23 ◽

Cited By ~ 10

Author(s):

Juliana Salazar ◽

Raimon Ferré ◽

Joan-Carles Vallvé ◽

Miguel Pocovı́ ◽

Manuel Castro Cabezas ◽

...

Keyword(s):

Retinoic Acid ◽

Single Nucleotide Polymorphisms ◽

Binding Protein ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

Acid Binding Protein ◽

Crabp Ii

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Direct Channeling of Retinoic Acid between Cellular Retinoic Acid-Binding Protein II and Retinoic Acid Receptor Sensitizes Mammary Carcinoma Cells to Retinoic Acid-Induced Growth Arrest

Molecular and Cellular Biology ◽

10.1128/mcb.22.8.2632-2641.2002 ◽

2002 ◽

Vol 22 (8) ◽

pp. 2632-2641 ◽

Cited By ~ 196

Author(s):

Anuradha S. Budhu ◽

Noa Noy

Keyword(s):

Retinoic Acid ◽

Mammary Carcinoma ◽

Transcriptional Activity ◽

Retinoic Acid Receptor ◽

Binding Protein ◽

Carcinoma Cells ◽

Acid Receptor ◽

Acid Binding Protein ◽

Mammary Carcinoma Cells ◽

Crabp Ii

ABSTRACT Cellular retinoic acid-binding protein II (CRABP-II) is an intracellular lipid-binding protein that associates with retinoic acid with a subnanomolar affinity. We previously showed that CRABP-II enhances the transcriptional activity of the nuclear receptor with which it shares a common ligand, namely, the retinoic acid receptor (RAR), and we suggested that it may act by delivering retinoic acid to this receptor. Here, the mechanisms underlying the effects of CRABP-II on the transcriptional activity of RAR and the functional consequences of these effects were studied. We show that CRABP-II, a predominantly cytosolic protein, massively undergoes nuclear localization upon binding of retinoic acid; that it interacts with RAR in a ligand-dependent fashion; and that, in the presence of retinoic acid, the CRABP-II-RAR complex is a short-lived intermediate. The data establish that potentiation of the transcriptional activity of RAR stems directly from the ability of CRABP-II to channel retinoic acid to the receptor. We demonstrate further that overexpression of CRABP-II in MCF-7 mammary carcinoma cells dramatically enhances their sensitivity to retinoic acid-induced growth inhibition. Conversely, diminished expression of CRABP-II renders these cells retinoic acid resistant. Taken together, the data unequivocally establish the function of CRABP-II in modulating the RAR-mediated biological activities of retinoic acid.

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