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Identification of the catalytic site of rat liver glutathione peroxidase as selenocysteine
Abstract
A procedure was developed to isolate 75Se-labeled rat liver glutathione peroxidase (glutathione:H2O2 oxidoreductase, EC 1.11.1.9) at 30--50% purity with 20--30% yields in 4--5 days. Using these preparations of glutathione peroxidase, the selenium moiety in the enzyme was identified as selenocysteine by derivatizing the seleno group with either iodoacetate or ethylenimine in the intact protein, hydrolyzing the protein with 6 N HCl, and cochromatographing the 75Se-labeled products with known standards. Techniques employed were anion-exchange chromatography, cation-exchange chromatography, gel-permeation chromatography, two-dimensional thin-layer chromatography, and automated amino acid analysis. The selenocysteine moiety was identified as the catalytic site in glutathione peroxidase by specifically labeling the enzyme with [14C]iodoacetate on the 75Se-labeled selenium atom and fractionating the 14C, 75Se-labeled derivative after acid hydrolysis. It was concluded that the reduced form of glutathione peroxidase contains the selenocysteine selenol (-SeH) at the catalytic site.
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Cited by
Organoselenium and Organotellurium Compounds: Toxicology and Pharmacology
Nogueira, C. W., Zeni, G., & Rocha, J. B. T. (2004) · Chemical Reviews · doi:10.1021/cr0406559 · Cited by 1,926
Biochemistry of Oxidative Stress
Sies, H. (1986) · Angewandte Chemie International Edition in English · doi:10.1002/anie.198610581 · Cited by 1,306
Maes, M., Galecki, P., Chang, Y. S., & Berk, M. (2011) · Progress in Neuro-Psychopharmacology and Biological Psychiatry · doi:10.1016/j.pnpbp.2010.05.004 · Cited by 1,140
Toxicology and pharmacology of selenium: emphasis on synthetic organoselenium compounds
Nogueira, C. W., & Rocha, J. B. T. (2011) · Archives of Toxicology · doi:10.1007/s00204-011-0720-3 · Cited by 477
Ebselen, a selenoorganic compound as glutathione peroxidase mimic
Sies, H. (1993) · Free Radical Biology and Medicine · doi:10.1016/0891-5849(93)90028-s · Cited by 431
Metadata
- Authors
- Forstrom, J. W., Zakowski, J. J., & Tappel, A. L.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1978-06-27
- Volume
- 17
- Issue
- 13
- Pages
- 2639-2644
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W1977651412
- Cited by
- 472
- References
- 0
- FWCI
- 15.72
- Type
- journal article
- Language
- en
- Open access
- closed
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- CoChat. (2026). Citation check: Identification of the catalytic site of rat liver glutathione peroxidase as selenocysteine. https://app.cochat.ai/papers/10.1021/bi00606a028
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- Forstrom, J. W., Zakowski, J. J., & Tappel, A. L. (1978). Identification of the catalytic site of rat liver glutathione peroxidase as selenocysteine. Biochemistry. https://doi.org/10.1021/bi00606a028
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