Recombinant human retinoic acid receptor α

Keidel, Siegfried; Rupp, Eva; Szardenings, Michael

doi:10.1111/j.1432-1033.1992.tb16739.x

Cited by 16 publications

(21 citation statements)

References 46 publications

(26 reference statements)

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“…The equilibrium dissociation constants (Kd; see Fig. 2 legend) obtained for RAR/pATH fusion proteins are in the range of values reported for RARs expressed in other prokaryotic (6,27,28) and eukaryotic expression systems (3,23,29,30 tors, and was only 4-to 7-fold less potent than authentic all-trans-RA. All-trans-retinol and 9-cis-RA were nearly equipotent in binding to the three subtypes of receptors.…”

Section: Competition Ofmentioning

confidence: 51%

All-trans-retinol is a ligand for the retinoic acid receptors.

Repa

Hanson

Clagett‐Dame

1993

Proc. Natl. Acad. Sci. U.S.A.

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Competition of all-trans-retinol and all-trans-retinaldehyde with 3H-labeled all-trans-retinoic acid (RA) for binding to retinoic acid receptors (RARs) was examined in human neuroblastoma cell nuclear extracts. All-trans-retinol was 35-fold less potent than all-trans-RA, whereas all-trans-retinaldehyde was 500-fold less active in binding to the nuclear receptors. To confirm that all-trans-retinol binds to RARs, experiments were carried out with RARs alpha, beta, and gamma expressed as bacterial fusion proteins. All-trans-retinol was only 4- to 7-fold less potent than all-trans-RA in binding to all three RAR subtypes. The all-trans-retinol binding observed was not the result of metabolism of retinol to RA or some other active compound during the binding experiment. Retinyl acetate was virtually inactive in competition binding experiments, while very slight activity was observed with 13-cis-RA and all-trans-retinaldehyde. Significant competition occurred with 4-hydroxy-RA and 4-keto-RA, which were 15- to 40-fold less potent than all-trans-RA. The 9-cis isomer of RA was equipotent with all-trans-retinol in these studies. These results suggest that all-trans-retinol cannot be excluded as a physiologically significant ligand for RAR-mediated gene expression.

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Section: Competition Ofmentioning

confidence: 51%

All-trans-retinol is a ligand for the retinoic acid receptors.

Repa

Hanson

Clagett‐Dame

1993

Proc. Natl. Acad. Sci. U.S.A.

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“…In addition to recently published results with human rRARa (Keidel et al, 1992), we can now discuss an even larger number of retinoids and their binding affinities for both proteins. All investigated retinoids, expect TTAB in case of rRARa, exhibit lower affinity for rCRABP-I and rRARa than retinoic acid.…”

Section: Retinoidmentioning

confidence: 99%

“…6; for rRARa, see Keidel et al, 1992). The natural retinoids retinal and retinol hardly competed, and ligands for other nuclear recptors did not compete with retinoic acid (Keidel et al, 1992).…”

Section: Binding Of the Retinoids To Recombinant (R) Crabp-i And Rraramentioning

confidence: 99%

“…6, the dose/response curves for inhibition of specific binding of [3H]retinoic acid to rCRABP-I by retinal, retinol and several synthetic retinoids are compared to that of retinoic acid (for rRARa, see Keidel et al, 1992). The binding of the investigated synthetic retinoids to rCRABP-I and rRARa is compared to their ability to induce alkaline phosphatase in neonatal rat heart cells, P19 teratocarcinoma cells and human prostate carcinoma cells (Table 1).…”

Section: Binding Of the Retinoids To Recombinant (R) Crabp-i And Rraramentioning

confidence: 99%

“…The affinities of the investigated retinoids to murine rCRABP-I (Zhang et al, 1992a) and human rRARa (Keidel et al, 1992) were determined using the charcoal-absorption hormone-binding assay with y-globulin instead of gelatin as expander protein (Keidel et al, 1992). The specifity of retinoic-acid binding was controlled by analysing [3H]retinoicacid-labelled bacterial extracts by sucrose-density-gradient centrifugation.…”

Section: Binding Of the Retinoids To Recombinant (R) Crabp-i And Rraramentioning

confidence: 99%

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In vivo biological activity of retinoids partially correlates to their affinity to recombinant retinoic‐acid receptor α and recombinant‐cellular retinoic‐acid‐binding protein I

Keidel

Szardenings

Mueller

1993

European Journal of Biochemistry

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Several known and some new retinoids were synthesized and their in vivo activity was investigated by an assay, based on induction of alkaline phosphatase in P19 teratocarcinoma cells, human prostate carcinoma cells and primary cultures of neonatal rat heart cells. The assay used in this study was found to be reproducible and useful for rapid screening of retinoids for biological activity. Two newly synthesized compounds exhibit high biological activity. The biological potency of the compounds was compared to their ability to bind to recombinant retinoic-acid receptor a and to cellular retinoic-acid-binding protein I determined by Charsorb-binding assay. mRNA of both retinoic-acid-binding proteins could be detected in the three cell lines investigated. As expected from the number of different retinoic-acid receptors, the results suggest that retinoids do not need to bind retinoic-acid receptor a nor cellular retinoic-acid-binding protein I in order to exhibit biological activity, but most retinoids investigated show a clear correlation between binding to these proteins and their biological activity.

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Characterization of heterologously expressed recombinant retinoic acid receptors with natural or synthetic retinoids

Kim

Sharma

1994

J. Biochem. Toxicol.

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The first step in retinoid action is binding to their nuclear receptors. Therefore, characterization of binding characteristics of retinoids is of major importance. Human retinoic acid receptors alpha (hRAR alpha), hRAR beta, and mouse RAR gamma (mRAR gamma) were expressed heterologously in Escherichia coli as a recombinant glutathione S-transferase (GST) fusion protein. The expressed fusion proteins were functional and bound specifically to the all-trans-retinoic acid (RA). The dissociation constants (Kd) for RA were 1.4 nM for GST-hRAR alpha, 1.4 nM for GST-hRAR beta, and 3.3 nM for GST-mRAR gamma, respectively. The fusion proteins were further used for competitive displacement assays to determine the displacement constant (DC50) for other selected retinoids. All-trans-RA and 4-oxo-all-trans-RA have high affinity with all three receptors (DC50 = 0.8-55 nM). The 13-cis RA binds to hRAR alpha with low affinity, but not to other RARs evaluated here. All-trans-N-ethylretinamide, all-trans-retinylacetate, and an ethyl ester of tetrahydronaphthalene derivative had no affinity to any RARs. The hRAR alpha and mRAR gamma receptors did not bind a naphthalene carboxylic acid derivative of RA, but hRAR beta binds this chemical with high affinity. Results indicated that the three recombinant proteins were functional in binding various RA congeners. The affinity and binding data of these retinoids were compared to their observed teratogenic activity.

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Recombinant human retinoic acid receptor α

Cited by 16 publications

References 46 publications

All-trans-retinol is a ligand for the retinoic acid receptors.

All-trans-retinol is a ligand for the retinoic acid receptors.

In vivo biological activity of retinoids partially correlates to their affinity to recombinant retinoic‐acid receptor α and recombinant‐cellular retinoic‐acid‐binding protein I

Characterization of heterologously expressed recombinant retinoic acid receptors with natural or synthetic retinoids

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