Solubilized TSH-receptor: Its Usefulness for the Radioligand Receptor Assay for TSH and TSH-displacing Antibody

Overview

Journal J Endocrinol Invest

Publisher Springer

Specialty Endocrinology

Date 1981 Apr 1

PMID 6268690

Citations 2

Authors

P Kotulla

H Schleusener

Affiliations

Soon will be listed here.

Abstract

The usefulness of solubilized TSH-receptor for measuring TSH and TSH-displacing antibodies (TDA) in a radioligand receptor assay (RRA) was investigated. Crude thyroid membranes were solubilized with Triton X-100. A soluble protein fraction was isolated by DEAE-Sephadex A-50 ion-exchange chromatography. The binding of 125I-TSH to the solubilized fraction was studied. It was maximal after 45 min at 37 C, whereas at 4 C a similar binding was observed after at least 17 h. The binding was inhibited by Ca2+ and Mg2+ with concentrations higher than 1 mM and by NaCl in the concentration range 17-150 mM. No inhibition of binding was found with normal serum up to 100 microliter. In contrast, 1.5 microliter of a Graves' disease serum showed significant inhibition. Using the system to measure TSH in serum, as low as 10 microunits TSH could be estimated. Compared to an RRA using thyroid membrane fractions, the sensitivity for TSH in serum as well as for TDA is increased at least tenfold in our system.

Citing Articles

TSH-displacing activity versus TSH-binding inhibiting activity of immunoglobulins from patients with Graves' disease.

Macchia E, Fenzi G, Monzani F, Bartalena L, Lippi F, Aloisio V J Endocrinol Invest. 1983; 6(5):375-8.

PMID: 6141201 DOI: 10.1007/BF03347619.

Radioligand receptor assay for TSH and TSH-binding-inhibiting antibodies using solubilized TSH-receptor.

Kotulla P, Kostner C, Schleusener H J Endocrinol Invest. 1983; 6(2):135-8.

PMID: 6134766 DOI: 10.1007/BF03350586.

References

AMIR S, Goldfine I, Ingbar S . Properties of the interaction between bovine thyrotropin and bovine thyroid plasma membranes. J Biol Chem. 1976; 251(15):4693-9. View

Endo K, Kasagi K, Konishi J, Ikekubo K, Okuno T, Takeda Y . Detection and properties of TSH-binding inhibitor immunoglobulins in patients with Graves' disease and Hashimoto's thyroiditis. J Clin Endocrinol Metab. 1978; 46(5):734-9. DOI: 10.1210/jcem-46-5-734. View

Tate R, Holmes J, Kohn L, Winand R . Characteristics of a solubilized thyrotropin receptor from bovine thyroid plasma membranes. J Biol Chem. 1975; 250(16):6527-33. View

ODonnell J, Trokoudes K, Silverberg J, Row V, Volpe R . Thyrotropin displacement activity of serum immunoglobulins from patients with Graves' disease. J Clin Endocrinol Metab. 1978; 46(5):770-7. DOI: 10.1210/jcem-46-5-770. View

Fenzi G, Macchia E, Bartalena L, Mazzanti F, BASCHIERI L, DeGroot L . Radio-receptor assay of TSH: its use to detect thyroid-stimulating immunoglobulins. J Endocrinol Invest. 1978; 1(1):17-24. DOI: 10.1007/BF03346766. View

Smith B, Hall R . Thyroid-stimulating immunoglobulins in Graves' disease. Lancet. 1974; 2(7878):427-31. DOI: 10.1016/s0140-6736(74)91815-7. View

Tate R, Schwartz H, Holmes J, Kohn L . Thyrotropin receptors in thyroid plasma membranes. Characteristics of thyrotropin binding and solubilization of thyrotropin receptor activity by tryptic digestion. J Biol Chem. 1975; 250(16):6509-15. View

Cuatrecasas P . Properties of the insulin receptor isolated from liver and fat cell membranes. J Biol Chem. 1972; 247(7):1980-91. View

Lee G, Aloj S, Beguinot F, Kohn L . Existence of a soluble thyrotropin binding component in normal human sera. J Biol Chem. 1977; 252(22):7967-70. View

10.

Smith B . Immunology of the thyrotropin receptor. Immunol Commun. 1976; 5(5):345-60. DOI: 10.3109/08820137609033853. View

11.

Schleusener H, Kotulla P, Finke R, Sorje H, Meinhold H, Adlkofer F . Relationship between thyroid status and Graves' disease-specific immunoglobulins. J Clin Endocrinol Metab. 1978; 47(2):379-84. DOI: 10.1210/jcem-47-2-379. View

12.

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

13.

Bashford C, Harrison S, Radda G . The relation between lipid mobility and the specific hormone binding of thyroid membranes. Biochem J. 1975; 146(2):473-9. PMC: 1165326. DOI: 10.1042/bj1460473. View

14.

Dawes P, Petersen V, Smith B, Hall R . Solubilization and partial characterization of human and porcine thyrotrophin receptors. J Endocrinol. 1978; 78(1):89-102. DOI: 10.1677/joe.0.0780089. View

15.

Manley S, BOURKE J, Hawker R . The thyrotrophin receptor in guinea-pig thyroid homogenate: general properties. J Endocrinol. 1974; 61(3):419-36. DOI: 10.1677/joe.0.0610419. View