Nature of steroid-glucocorticoid receptor interactions: thermodynamic analysis of the binding reaction

Wolff, Manfred E.; Baxter, John D.; Kollman, Peter A.; Lee, David L.; Kuntz, Irwin D.; Bloom, Ernest; Matulich, Daniel T.; Morris, Julie

doi:10.1021/bi00609a005

Cited by 104 publications

(33 citation statements)

References 42 publications

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“…It is known that hydrophobic interactions are important for high receptor binding affinity of steroids, whereas hydrogen bonds might provide ligand binding specificity (40). Mutagenesis of Met-560 to leucine did not affect binding affinity for any of the tested ligands (Table I and Fig.…”

Section: Discussionmentioning

confidence: 92%

Functional Probing of the Human Glucocorticoid Receptor Steroid-interacting Surface by Site-directed Mutagenesis

Lind

Greenidge

Gillner

et al. 2000

Journal of Biological Chemistry

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To elucidate which amino acids in the glucocorticoid receptor ligand-binding domain might be involved in determining steroid binding specificity by interaction with the D-ring of glucocorticoids, we have performed site-directed mutagenesis of the four amino acids Met-560, Met-639, Gln-642, and Thr-739 based on their proximity to the steroid in a model structure. Mutations of these residues affected steroid binding affinity, specificity, and/or steroid-dependent transactivation. The results indicate that these residues are located in close proximity to the ligand and appear to play a role in steroid recognition and/or transactivating sensitivity, possibly by changes in the steroid-dependent conformational change of this region, resulting in the formation of the AF-2 site. Mutation of Gln-642 resulted in a marked decrease in affinity for steroids containing a 17␣-OH group. This effect was alleviated by the presence of a 16␣-CH 3 group to a varying degree. Thr-739 appears to form a hydrogen bond with the 21-OH group of the steroid, as well as possibly forming hydrophobic interactions with the steroid. Met-560 and Met-639 appear to form hydrophobic interactions with the D-ring of the steroid, although the nature of these interactions cannot be characterized in more detail at this point. The glucocorticoid receptor (GR)1 belongs to the superfamily of hormone-dependent nuclear receptors and consists of three structural and functional main domains: the N-terminal domain, which harbors the major transactivating function (AF-1); the central domain, which binds to DNA in glucocorticoid regulated genes; and the C-terminal domain, which binds the ligand (1-4).The ligand-binding domain (LBD) comprises approximately 250 amino acids and is in its unliganded state associated with a complex containing heat shock proteins and immunophilins (5). Upon ligand binding this complex dissociates and a cascade of events are triggered leading to induction or repression of target genes. Within the ligand-binding domain there are also a hormone-dependent nuclear localization signal (6) and hormone-dependent transactivation functions (AF-2) (7-10).The crystal structure of the GR LBD is not yet available, but the crystal structures of the LBDs of other members of the nuclear receptor superfamily including the peroxisome proliferator activated receptor, retinoic acid receptor, retinoid X receptor, thyroid hormone receptor, progesterone receptor (PR), estrogen receptor ␣ (ER␣), and estrogen receptor ␤ (ER␤) have been solved (11-17). Their structures contain 12 ␣-helices that are folded in a very similar way into a three-layered antiparallel ␣-helical sandwich that creates a hydrophobic pocket for the ligand. Upon ligand binding a conformational change occurs, mainly involving helix 12, which folds up against the protein body and creates a lid for the ligand binding pocket. This also leads to formation of the AF-2 interface, which has been shown to interact with transcriptional coactivators (18 -21).The mechanisms that determine the binding affi...

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Section: Discussionmentioning

confidence: 92%

Functional Probing of the Human Glucocorticoid Receptor Steroid-interacting Surface by Site-directed Mutagenesis

Lind

Greenidge

Gillner

et al. 2000

Journal of Biological Chemistry

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“…Rats were weighed, anesthetized with ketamine/xylazine, and sacrificed by aortic exsanguinations at 0.25, 0.5, 0.75, 1, 2, 4, 5,6,7,8,12,24,36,48,60,72,84 and 96 h (n = 3 per time point). Six rats injected with IM saline and sacrificed at 12 and 24 h (n = 3 per time point) served as controls.…”

Section: Methodsmentioning

confidence: 99%

“…10) the dynamics of which are governed by MPL, to form drug-receptor complex in the cytosol (CR), which rapidly translocates to the nucleus forming DR(N). Both nuclear drug-receptor complexes act non-competitively on TAT mRNA production rate as given by, (18) (19) (20) where K d,CST is the dissociation constant for CST binding to free receptors (R) and was calculated from a quantitative structure-property relationship (QSPR) model [36]; k T is the translocation constant to account for rapid entry of CR to form CR(N). The TAT mRNA is produced at a zero-order rate (k s,TATm ) and dissipates with a first-order rate constant, k d,TATm .…”

Section: Tyrosine Aminotransferase Dynamicsmentioning

confidence: 99%

Modeling receptor/gene-mediated effects of corticosteroids on hepatic tyrosine aminotransferase dynamics in rats: dual regulation by endogenous and exogenous corticosteroids

Hazra

Pyszczynski

DuBois

et al. 2007

J Pharmacokinet Pharmacodyn

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Receptor/gene-mediated effects of corticosteroids on hepatic tyrosine aminotransferase (TAT) were evaluated in normal rats. A group of normal male Wistar rats were injected with 50 mg/kg methylprednisolone (MPL) intramuscularly at the nadir of their plasma corticosterone (CST) rhythm (early light cycle) and sacrificed at various time points up to 96 h post-treatment. Blood and livers were collected to measure plasma MPL, CST, hepatic glucocorticoid receptor (GR) mRNA, cytosolic GR density, TAT mRNA, and TAT activity. The pharmacokinetics of MPL showed bi-exponential disposition with two first-order absorption components from the injection NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript site and bioavailability was 21%. Plasma CST was reduced after MPL dosing, but resumed its daily circadian pattern within 36 h. Cytosolic receptor density was significantly suppressed (90%) and returned to baseline by 72 h resuming its biphasic pattern. Hepatic GR mRNA follows a circadian pattern which was disrupted by MPL and did not return during the study. MPL caused significant down-regulation (50%) in GR mRNA which was followed by a delayed rebound phase (60-70 h). Hepatic TAT mRNA and activity showed up-regulation as a consequence of MPL, and returned to their circadian baseline within 72 and 24 h of treatment. A mechanistic receptor/genemediated pharmacokinetic/pharmacodynamic model was able to satisfactorily describe the complex interplay of exogenous and endogenous corticosteroid effects on hepatic GR mRNA, cytosolic free GR, TAT mRNA, and TAT activity in normal rats.

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“…(Ross & Subramanian, 1981;Strazza et al, 1985). The fact that the sign of AS$,.Oc' is positive is further evidence that the driving forces for the formation of the transition state are hydrophobic; a negative value would be expected if there were more hydrogen bonds in the transition state than in the reactants (Wolff et al, 1978). It appears that the first event is most appropriately described in terms of the 'hydrophobically bound complex' (terminology of Ross & Subramanian, 1981).…”

Section: Free Energy Enthalpy Entropy Of Prl-receptor Reactionmentioning

confidence: 96%

“…Changes in secondary and tertiary structure accompanying the association resulted in the curvilinear van't Hoff plot and free-energy change (Osborne et al, 1976). Over a broader temperature range, curvilinear van't Hoff plots or free-energy changes have been reported for the follitropin receptor (Andersen et al, 1983;Sanborn et al, 1987), insulin receptor (Waelbroeck et al, 1979), glucocorticoid receptor (Wolff et al, 1978), and self-associating proteins (Osborne et al, 1976;Formisano et al, 1977;Ross & Subramanian, 1981). In the present study the temperature varied within a relatively narrow range, and the effects of temperature Table 1.…”

Section: Free Energy Enthalpy Entropy Of Prl-receptor Reactionmentioning

confidence: 99%

Thermodynamic analysis of the interaction of prolactin with its receptor in the rabbit mammary-gland microsomes

Sakai

Suzuki

Kohmoto

1990

Biochemical Journal

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The interaction of prolactin (PRL) with its membrane receptor depends markedly on temperature. Thermodynamic parameters for this reaction have been-evaluated from data for time-course kinetics and equilibrium binding at multiple temperatures between 19 and 31 'C. The free-energy change with temperature and the van't Hoff plot were found to be linear. These suggest that there are minimal structure changes at the PRL-receptor contact site over this temperature range. The positive signs of the entropy and enthalpy of reaction, and of the entropy of activation (AST) for association, indicate that the hydrophobic bonding is the most significant force involved in PRL-receptor formation. The AS-for dissociation was negative, and the enthalpy of activation for dissociation was about 20.3 kJ mol-' larger than that for association, indicating that the PRL-receptor complex is further stabilized by contributions of hydrogen bonds and van der Waals contacts after the initial interaction. The free energy of activation for dissociation, at 25 'C, was about 2.5-fold larger than that for association. This would cause slow dissociation of PRL from its receptor.

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Nature of steroid-glucocorticoid receptor interactions: thermodynamic analysis of the binding reaction

Cited by 104 publications

References 42 publications

Functional Probing of the Human Glucocorticoid Receptor Steroid-interacting Surface by Site-directed Mutagenesis

Functional Probing of the Human Glucocorticoid Receptor Steroid-interacting Surface by Site-directed Mutagenesis

Modeling receptor/gene-mediated effects of corticosteroids on hepatic tyrosine aminotransferase dynamics in rats: dual regulation by endogenous and exogenous corticosteroids

Thermodynamic analysis of the interaction of prolactin with its receptor in the rabbit mammary-gland microsomes

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