Total and high-affinity corticosteroid-binding globulin depletion in septic shock is associated with mortality

2018 ◽  
Vol 90 (1) ◽  
pp. 232-240 ◽  
Author(s):  
Emily J. Meyer ◽  
Marni A. Nenke ◽  
Wayne Rankin ◽  
John G. Lewis ◽  
Elisabeth Konings ◽  
...  
Keyword(s):  
Septic Shock ◽  
High Affinity ◽  
Corticosteroid Binding Globulin

scholarly journals OR19-05 Steroid:Corticosteroid-Binding Globulin Interactions; Effects of Neutrophil Elastase Cleavage, Pyrexia and Acidosis

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Emily Jane Meyer ◽  
David J Torpy ◽  
Anastasia Chernykh ◽  
Morten Thaysen-Anderson ◽  
Marni Anne Nenke ◽  
...  
Keyword(s):  
Septic Shock ◽  
Binding Affinity ◽  
Neutrophil Elastase ◽  
Binding Constants ◽  
Hormone Binding ◽  
Binding Characteristics ◽  
High Affinity ◽  
Free Cortisol ◽  
Corticosteroid Binding Globulin ◽  
17 Hydroxyprogesterone

Abstract Context Corticosteroid-binding globulin (CBG) transports cortisol and other steroid hormones1,2. High-affinity CBG (haCBG) undergoes proteolysis of the reactive centre loop (RCL) by neutrophil elastase (NE) at inflammatory sites, liberating immunomodulatory cortisol and altering conformation to low-affinity CBG (laCBG). Pyrexia reduces CBG:cortisol binding affinity, an interaction at the RCL is speculated3. Objective To measure the equilibrium binding constants of a panel of steroids to glycosylated haCBG and laCBG over temperature and pH ranges mimicking the pathophysiological conditions of septic shock. Design Surface plasmon resonance was used to determine the binding profiles of 19 steroid ligands to haCBG and laCBG at temperatures 25°C, 37°C and 39°C and pH 7.4 and 7.0. The RCL-recognizing 9G12 antibody was used to assess cleavage and epitope availability of the RCL across conditions. Results A 4–8 fold reduction in affinity for cortisol, cortisone, corticosterone, 11-deoxycortisol, progesterone, 17-hydroxyprogesterone and prednisolone occurred with NE-mediated haCBG-to-laCBG conversion, cortisol expectedly displayed the highest binding affinity. Binding affinity consistently decreased at higher temperatures and at acidic pH for both haCBG and laCBG. 9G12 antibody RCL binding was preserved for haCBG across temperatures. Conclusions These studies reveal that steroid binding to CBG is selective and in all cases reduced upon NE-mediated haCBG-to-laCBG transition. Moreover, reduced CBG:cortisol binding affinity at elevated temperature occurs with an intact and accessible RCL epitope, suggesting a non-RCL mechanism for the delivery of anti-inflammatory cortisol in pyrexia. Synergy of NE cleavage and pyrexia/acidosis may serve for local inflammatory site cortisol delivery and increase free cortisol. These findings demonstrate the modifiable hormone binding characteristics of CBG in (patho-)physiological conditions, supporting its significance in cortisol delivery in obviating systemic inflammation and multiorgan-organ failure in patients with septic shock and its association with mortality4. 1. Pemberton PA, Stein PE, Pepys MB, et al. Hormone binding globulins undergo serpin conformational change in inflammation. Nature. 1988;336(6196):257–258. 2. Pugeat MM, Dunn JF, Nisula BC. Transport of steroid hormones: interaction of 70 drugs with testosterone-binding globulin and corticosteroid-binding globulin in human plasma. J Clin Endocrinol Metab. 1981;53(1):69–75. 3. Cameron A, Henley D, Carrell R, et al. Temperature-responsive release of cortisol from its binding globulin: a protein thermocouple. J Clin Endocrinol Metab. 2010;95(10):4689–4695. 4. Meyer EJ, Nenke MA, Rankin W, et al. Total and high-affinity corticosteroid-binding globulin depletion in septic shock is associated with mortality. Clin Endocrinol (Oxf). 2019;90(1):232–240.

CHARACTERIZATION AND ASSAY OF THE PROGESTERONE RECEPTOR IN RAT UTERINE CYTOSOL

1978 ◽  
Vol 76 (1) ◽  
pp. 21-31 ◽  
Author(s):  
M. T. VU HAI ◽  
E. MILGROM
Keyword(s):  
Progesterone Receptor ◽  
Receptor Complexes ◽  
High Affinity ◽  
Receptor Sites ◽  
Natural Hormone ◽  
Corticosteroid Binding Globulin ◽  
Steroid Binding Proteins ◽  
Steroid Binding ◽  
High Concentrations ◽  
Synthetic Progestogen

SUMMARY The synthetic progestogen R5020 (17,21-dimethyl-19-norpregna-4,9-diene-3,20-dione) binds with high affinity (Ka = 8·8 × 108 1/mol at 0 °C) to the progesterone receptor from rat uterine cytosol. At nanomolar concentrations, equilibrium is attained in less than 90 min. R5020 has a very low affinity for other specific steroid-binding proteins (corticosteroid-binding globulin and oestrogen receptors) present in relatively high concentrations in the uterine cytosol. The affinity of the receptor for the natural hormone progesterone is remarkably low (Ka= 1 × 108−1·7 × 1081/mol at 0 °C) which explains the instability of progesterone–receptor complexes. Advantage may be taken of this property to remove endogenous progesterone easily by charcoal treatment at 0 °C, a treatment which does not modify the concentration of receptors. A method based on these characteristics is described for the assay of the total number (progesterone-bound and unbound) of receptor sites in uterine cytosol. This assay may be used in various physiological situations where endogenous progesterone is present at unknown concentrations.

Binding of 5α-dihydroprogesterone to proteins in human pregnancy serum

1982 ◽  
Vol 100 (1) ◽  
pp. 120-130
Author(s):  
Oscar A. Lea
Keyword(s):  
Protein Separation ◽  
Specific Binding ◽  
Sex Hormone Binding Globulin ◽  
Complex Stability ◽  
Hormone Binding ◽  
Acid Glycoprotein ◽  
High Affinity ◽  
Corticosteroid Binding Globulin ◽  
In Pregnancy ◽  
Binding Component

Abstract. The binding of 5α-dihydroprogesterone (DHP)1 to proteins in pregnancy serum has been investigated and compared with the binding of progesterone. Characteristic properties of the DHP-serum protein interaction were unsaturability, low affinity and poor complex stability. Fractionation of serum using a variety of protein separation techniques, revealed that DHP interacts with several proteins. At low temperature (0–4°C) albumin appeared to be the principal binding component whereas higher temperatures seemed to favour binding to β-lipoproteins and α2-macroglobulin. Binding to specific binding proteins such as the corticosteroid binding globulin (CBG) and the sex hormone-binding globulin, (SHBG) were detectable but appeared to be quantitatively unimportant. Progesterone showed a similar multicomponent interaction but differed from DHP in the extent of binding to CBG. Binding of either hormone to the α1-acid-glycoprotein was negligible. The present study shows that the high endogenous DHP levels present in pregnancy sera are caused by factors other than high affinity protein binding.

Decreased immunoreactivity and binding activity of corticosteroid-binding globulin in serum in septic shock.

1989 ◽  
Vol 35 (8) ◽  
pp. 1675-1679 ◽  
Author(s):  
M Pugeat ◽  
A Bonneton ◽  
D Perrot ◽  
B Rocle-Nicolas ◽  
H Lejeune ◽  
...  
Keyword(s):  
Septic Shock ◽  
Ion Exchange ◽  
Binding Capacity ◽  
Normal Subjects ◽  
Binding Activity ◽  
Healthy Controls ◽  
Corticosteroid Binding Globulin ◽  
Ion Exchange Column ◽  
Monospecific Antiserum ◽  
Final Purification

Abstract To investigate the mechanism(s) responsible for the depletion of corticosteroid-binding globulin (CBG) activity in serum in septic shock, we developed a radioimmunoassay (RIA) for human CBG, using a monospecific antiserum to human CBG raised in rabbits. CBG was purified from pooled human serum by precipitation with ammonium sulfate and successive affinity chromatography treatments on corticosterone-Sepharose and concanavalin A-Sepharose. Final purification was achieved by HPLC on a diethylaminoethyl-PW (polymer matrix) ion-exchange column. Typical standard curves established for the CBG immunoassay showed parallelism for pure CBG and serial dilutions of sera from patients with septic or nonseptic shock and from healthy controls. Measurements of CBG by RIA showed a significantly (P less than 0.001) lower CBG concentration in patients with septic shock (22.9 +/- 5.9 mg/L, mean +/- SD; n = 23) than in controls (39.9 +/- 6.5 mg/L, n = 21) or in patients with nonseptic shock (33.3 +/- 6.5 mg/L, n = 12). The correlation between the concentrations determined by RIA and the CBG binding capacity was significant (r = 0.619, P less than 0.001, n = 33). The electrophoretic mobility of CBG was similar in sera from septic shock patients and normal subjects (Rf = 0.52-0.56). This suggests that the depletion of the corticosteroid-binding activity in serum during septic shock is associated with a decreased amount of CBG.

A one-tube micromethod for radioimmunoassay of plasma cortisol.

1978 ◽  
Vol 24 (11) ◽  
pp. 1954-1957 ◽  
Author(s):  
G Ellis ◽  
R Morris
Keyword(s):  
Aqueous Phase ◽  
Plasma Cortisol ◽  
Sulfate Solution ◽  
Ammonium Sulfate Solution ◽  
Micro Scale ◽  
Standard Curve ◽  
Scale Method ◽  
High Affinity ◽  
Free Cortisol ◽  
Corticosteroid Binding Globulin

Abstract A micro-scale method is presented for measurement of cortisol. Serum or plasma, 2 microliter, is diluted with buffer and the solution heated at 60 degrees C in the presence of high-affinity antibody and [3H] cortisol. Corticosteroid-binding globulin in the serum is denatured, releasing cortisol and making it available to the antibody, which is stable during the incubation. After cooling, ammonium sulfate solution is added, followed by a toluene-based scintillant that does not dissolve in the aqueous phase. The vial is shaken to extract the free cortisol into the scintillant. Antibody bound cortisol remains in the aqueous phase and does not produce any scintillation. The amount of [3H] cortisol that is free at the end of the incubation, and which therefore gives rise to the measured counts, is related to the concentration of cortisol in the original serum, and may be calculated from a standard curve. The assay is sensitive and precise. Ranges are presented for normal and pathologic subjects.

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