Affinity purification of retinoic acid-binding proteins using immobilized 4-(2-hydroxyethoxy)retinoic acid.

  title={Affinity purification of retinoic acid-binding proteins using immobilized 4-(2-hydroxyethoxy)retinoic acid.},
  author={J. M. Jun. Chapman and R. W. Jun. Curley},
  journal={Protein expression and purification},
  volume={1 1},
  • J. Chapman, R. Curley
  • Published 1 September 1990
  • Biology, Medicine
  • Protein expression and purification
An affinity chromatographic matrix that purifies cellular retinoic acid-binding protein to near homogeneity from rat testes cytosol has been developed. The three-step procedure includes an acid precipitation, a batch treatment with CM Bio-Gel, and affinity chromatography on 4-(2-hydroxyethoxy)retinoic acid coupled to epoxy-activated Sepharose 6B. The binding protein was purified approximately 8500-fold based on total soluble testicular protein and with a recovery in excess of 80%. In addition… Expand
5 Citations
Protein knockdown using methyl bestatin-ligand hybrid molecules: design and synthesis of inducers of ubiquitination-mediated degradation of cellular retinoic acid-binding proteins.
4b possesses sufficient activity, permeability, and stability in cells to be employed in cellular assays and confirmed that 4b induced formation of a complex of cIAP1 and CRABP-II in vitro and induced proteasomal degradation of CRABP -II in cells. Expand
Retinoid Structure, Chemistry, and Biologically Active Derivatives
Abstract After a description of the structure and important physical properties of retinoic acid is completed, a brief overview of the metabolic processes leading to the production of retinoic acidExpand
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  • Medicine, Chemistry
  • Chemical & pharmaceutical bulletin
  • 2020
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Cellular retinoic acid-binding protein from rat testis. Purification and characterization.
  • D. Ong, F. Chytil
  • Chemistry, Medicine
  • The Journal of biological chemistry
  • 1978
Cellular retinoic acid-binding protein has been purified to homogeneity from rat testes, and the apparent dissociation constant was determined by fluorometric titration to be 4.2 X 10(-9) M. Expand
Affinity purification of cellular retinoic acid-binding protein on 14-carboxy-13-cis-retinamide-sepharose 4B.
The synthesis of 14-carboxy-13-cis-retinamide-Sepharose 4B is described, which is an affinity matrix bearing an all-trans-RA moiety, and thus was used to purify and characterize CRABP from chick-embryo skin and exhibited high efficiency and selectivity for RA-binding protein. Expand
Purification and partial characterization of a novel binding protein for retinoic acid from neonatal rat.
Amino-terminal sequence analysis showed that CRABP-II has a distinct sequence, while the CRA BP-I sequence is exactly identical to that of the rat testis CRABP. Expand
Retinoic acid-binding protein in rat tissue. Partial purification and comparison to rat tissue retinol-binding protein.
Tissue retinoic acid-binding protein can be detected in extracts of brain, eye, ovary, testis, and uterus but is apparently absent in heart muscle, small intestine, kidney, liver, lung, gastrocnemious muscle, serum, and spleen. Expand
Purification of retinol-binding protein from serum and urine by affinity chromatography.
A new method for the purification of retinol-binding protein from urine and serum is described, which has the same electrophoretic mobility and molecular weight and the same ability to interact with retinoic acid and prealbumin as retinols-bindingprotein prepared by conventional methods. Expand
Affinity chromatographic purification of serum retinol-binding protein using 4-substituted aminoretinoids.
Retinol-binding protein has been purified from rabbit serum by a new affinity chromatographic phase and employs a derivatized affinity ligand while preserving the integrity of the beta-ionone molecule via substitution of the allylic 4-position. Expand
Relationship between binding affinities to cellular retinoic acid-binding protein and in vivo and in vitro properties for 18 retinoids.
While all retinoids that are potent in these biological test systems bind tightly to cellular retinoic acid-binding protein, the converse is not true and the lack of a consistent quantitative correlation between 50% inhibitory concentration and biological activity is probably due to insufficient concentrations of the retinoid in the target tissue or celll. Expand
High pressure liquid chromatographic detection of intracellular retinoid binding proteins from cultured cell and tumor cytosols.
This method, which resolves CRABP and CRBP in crude mixtures from the majority of cytosolic proteins, should be a valuable tool in the evaluation of vitamin A-binding protein interactions and their biological significance. Expand
Localization of retinoic acid-binding protein in nuclei and the nuclear uptake of retinoic acid.
Retinoic acid-binding protein (RABP) has been detected in the nuclei of chick embryo skin and Lewis lung tumor. The nuclear binding component showed the same ligand specificity and sedimentationExpand
Occurrence of a binding protein for 11-cis-retinal in retina.
A binding protein for retinal has been found in the soluble protein fraction of bovine retina and is proposed to be referred to as 11-cis-retinal-binding protein, indicating that the functional group remained accessible when in the protein complex. Expand