Open AccessProstaglandin Synthetase System—Resolution into Oxygenase and Isomerase Components
Author(s) -
Tsumoru Miyamoto,
Shozo Yamamoto,
Osamu Hayaishi
Publication year - 1974
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.71.9.3645
Subject(s) - chemistry , prostaglandin , microsome , glutathione , chromatography , biochemistry , ethylene glycol , enzyme , stereochemistry , organic chemistry
The microsomal fraction of bovine vesicular gland catalyzed the conversion of [1 -14 C]8,11,14-eicosatrienoic acid to prostaglandin E1 in the presence of tryptophan, hemoglobin, and glutathione. The prostaglandin synthetase system was solubilized by treatment of the microsomal fraction with Tween 20 in the presence of ethylene glycol. DEAE-cellulose column chromatography separated the enzyme into two fractions (Fractions I and II), both of which were required for prostaglandin E1 synthesis. When Fraction I alone was incubated with 8,11,14-eicosatrienoic acid, an unstable compound accumulated. This compound was converted to prostaglandin E1 by the addition of Fraction II. On the basis of itsRF values on thin-layer chromatography, its reduction to prostaglandin F1α with stannous chloride, and the decomposition to prostaglandins F1α , E1 , and D1 at room temperature, the unstable intermediate was tentatively identified to be the 9,11-endoperoxide derivative, referred to previously as prostaglandin R1 (Nugterenet al. (1973)Biochim. Biophys. Acta 326, 448-461) or prostaglandin H1 (Hamberget al. (1974)Proc. Nat. Acad. Sci. USA 71, 345-349). Of the three cofactors mentioned above, Fraction I required both tryptophan and hemoglobin, while only glutathione was necessary for Fraction II. Anti-inflammatory agents such as indomethacin and aspirin were inhibitory to Fraction I.