Role of Regulatory F-domain in Hepatocyte Nuclear Factor-4α Ligand Specificity

Petrescu, Anca D.; Hertz, Rachel; Bar‐Tana, Jacob; Schroeder, Friedhelm; Kier, Ann B.

doi:10.1074/jbc.m405906200

Cited by 23 publications

(37 citation statements)

References 52 publications

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“…Interestingly, helix 12, but not the F domain, was found to be essential for ERa interactions with cytokeratins 8 and 18 (Long & Nephew 2006), suggesting an F domain-independent action that adds further complexity. Similar observations detailing the role of the F domain on the function of other NR proteins have been noted (Modarress et al 1997, Sladek et al 1999, Suaud et al 1999, Ruse et al 2002, Bertin et al 2004, Farboud & Privalsky 2004, Petrescu et al 2005, Takayama et al 2007). In particular, in a study of the hepatocyte nuclear factor-4a (HNF4A), the authors assert that the characteristics of F domain-truncated mutants 'may not reflect the structure, ligand-binding, ligand-conformational responsiveness and cooperativity of full-length HNF4A' (Petrescu et al 2005).…”

Section: Structural Validation Of a Second Nr Ligand-binding Sitesupporting

confidence: 75%

Implications of the binding of tamoxifen to the coactivator recognition site of the estrogen receptor

Kojetin¹,

Burris²,

Jensen³

et al. 2008

Endocrine Related Cancer

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A number of studies have reported on the unusual pharmacological behavior of type I antiestrogens, such as tamoxifen. These agents display mixed agonist/antagonist activity in a dose-, cell-, and tissue-specific manner. Consequently, many efforts have been made to develop so-called 'pure' antiestrogens that lack mixed agonist/antagonist activity. The recent report of the structure of estrogen receptor (ER) b with a second molecule of 4-hydroxytamoxifen (HT) bound in the coactivator-binding surface of the ligand-binding domain (LBD) represents the first direct example of a second ER ligand-binding site and provides insight into the possible origin of mixed agonist/antagonist activity of type I antiestrogens. In this review, we summarize the biological reports leading up to the structural conformation of a second ER ligand-binding site, compare the ERb LBD structure bound with two HT molecules to other ER structures, and discuss the potential for small molecular inhibitors designed to directly inhibit ER-coactivator and, more generally, nuclear receptor (NR)-coactivator interactions. The studies support a departure from the traditional paradigm of drug targeting to the ligand-binding site, to that of a rational approach targeting a functionally important surface, namely the NR coactivator-binding (activation function-2) surface. Furthermore, we provide evidence supporting a reevaluation of the strict interpretation of the agonist/antagonist state with respect to the position of helix 12 in the NR LBD.

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Section: Structural Validation Of a Second Nr Ligand-binding Sitesupporting

confidence: 75%

Implications of the binding of tamoxifen to the coactivator recognition site of the estrogen receptor

Kojetin¹,

Burris²,

Jensen³

et al. 2008

Endocrine Related Cancer

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“…Recombinant mouse ACBP with identical amino acid sequence as native ACBP was produced as described previously [4]. Full length hepatic nuclear factor-4α (HNF-4α) was expressed in E. coli and purified as previously described [29][30][31]37]. Cis-parinaric acid (naturally-fluorescent fatty acid analogue), was obtained from Molecular Probes, InVitrogen (Eugene, OR).…”

Section: Methodsmentioning

confidence: 99%

Structural and functional characterization of a new recombinant histidine-tagged acyl coenzyme A binding protein (ACBP) from mouse

Petrescu

Huang

Hostetler

et al. 2008

Protein Expression and Purification

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Acyl-coenzyme A binding protein (ACBP) has been proposed to transport fatty acyl-CoAs intracellularly, facilitating their metabolism. In this study, a new mouse recombinant ACBP was produced by insertion of a histidine (his) tag at the C-terminus to allow efficient purification by Niaffinity chromatography. The his-tag was inserted at the C-terminus since ACBP is a small molecular size (10 kDa) protein whose structure and activity are sensitive to amino acid substitutions in the Nterminus. The his tag had no or little effect on ACBP structure or ligand binding affinity and specificity. His-ACBP bound the naturally-occurring fluorescent cis-parinaroyl-CoA with very high affinity (K d =2.15 nM), but exhibited no affinity for non-esterified cis-parinaric acid. To determine if the presence of the C-terminal his tag altered ACBP interactions with other proteins, direct binding to hepatocyte nuclear factor 4α (HNF-4α), a nuclear receptor regulating transcription of genes involved in lipid metabolism, was examined. His-ACBP and HNF-4α were labeled with Cy5 and Cy3, respectively, and direct interaction was determined by a novel fluorescence resonance energy transfer (FRET) binding assay. FRET analysis showed that his-ACBP directly interacted with HNF-4α (intermolecular distance of 73 Å) at high affinity (K d =64-111 nM) similar to native ACBP. The his-tag also had no effect on ACBPs ability to interact with and stimulate microsomal enzymes utilizing or forming fatty acyl CoA. Thus, C-terminal his-tagged-ACBP maintained very similar structural and functional features of the untagged native protein and can be used in further in vitro experiments that require pure recombinant ACBP.

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“…Based on recent demonstrations that truncation of a nuclear transcription factor can signifi cantly affect ligandbinding affi nity, specifi city, and consequently, receptor activity ( 23,24 ), full-length hPPAR ␣ and mPPAR ␣ were mixed mPPAR ␣ and BODIPY C16-CoA at the same molar ratio used for the hPPAR ␣ displacement assays. However, very little displacement was noted for any ligand and only at high LCFA concentrations (data not shown).…”

Section: Full-length Hppar ␣ and Mppar ␣ Protein Purifi Cationmentioning

confidence: 99%

Divergence between human and murine peroxisome proliferator-activated receptor alpha ligand specificities

Oswal

Balanarasimha

Loyer

et al. 2013

Journal of Lipid Research

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Role of Regulatory F-domain in Hepatocyte Nuclear Factor-4α Ligand Specificity

Cited by 23 publications

References 52 publications

Implications of the binding of tamoxifen to the coactivator recognition site of the estrogen receptor

Implications of the binding of tamoxifen to the coactivator recognition site of the estrogen receptor

Structural and functional characterization of a new recombinant histidine-tagged acyl coenzyme A binding protein (ACBP) from mouse

Divergence between human and murine peroxisome proliferator-activated receptor alpha ligand specificities

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