Solubilization of thyroid peroxidase by nonionic detergents

• Published in: The Journal of biological chemistry Vol. 251; no. 8; pp. 2525 - 2529
• Main Authors: Neary, J T, Davidson, B, Armstrong, A, Strout, H V, Maloof, F
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 25.04.1976
• Subjects: Animals; Hydrogen-Ion Concentration; Peroxidases - isolation & purification; Peroxidases - metabolism; Polyethylene Glycols; Solubility; Structure-Activity Relationship; Surface-Active Agents; Swine; Thyroid Gland - enzymology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)33619-0

We have examined the ability of nonionic detergents to solubilize thyroid peroxidase from a porcine thyroid particulate fraction, as measured by the release of peroxidase activity into the supernatant fraction after centrifugation at 105,000 X g for 1 hour and the retardation of the supernatant peroxidase of Sepharose 6B. The parameters of peroxidase solubilization by Triton X-100 have been investigated in detail. Under optimum conditions, 60 to 95% of the thryoid peroxidase and about 50% of the total protein is released into the 105,000 X g, 1-hour supernatant. Under the optimum conditions established with Triton X-100, a series of Brij detergents of different chemical structure were equally effective in releasing peroxidase and protein. The protein patterns of the supernatants obtained with these detergents were similar on sodium dodecyl sulfate-polyacrylamide electrophoresis gels, suggesting that the detergents studied release similar membrane proteins. The Triton X-100 and Brij 58 supernatants were chromatographed separately on Sepharose 6B equilibrated with 0.1% Triton X-100 or Brij 58, respectively. In both cases, 75 to 80% of the peroxidase activity was retarded, thereby indicating that the nonionic detergents effect solubilization of the peroxidase rather than dispersal of nonsedimentable membrane fragments. These studies report the first successful solubilization of thyroid peroxidase by nonionic detergents. Together with previous evidence from our laboratory, these experiments indicate that thyroid peroxidase is an integral membrane protein.