Simple and rapid purification of tryptophan 5-monooxygenase from rabbit brain by affinity chromatography.
A simple and rapid method for isolating tryptophan 5-monooxygenase [L-tryptophan, tetrahydropteridine:oxygen oxidoreductase (5-hydroxylating), EC 1.14.16.4] was reported. The method involves adsorption on calcium phosphate gel and affinity chromatography on agarose coupled with dimethyltetrahydropteridine. Tryptophan 5-monooxygenase was purified 1,100-fold from a rabbit brain extract to a specific activity of 15.9 nmol/min.mg, which is far higher than that of the highly purified preparation reported by Tong and Kaufman ((1975) J. Biol.  Chem. 250, 4152-4158). It was also demonstrated that this method was applicable to the purification of rat brain tryptophan 5-monooxygenase.
D000327:Adsorption; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002130:Calcium Phosphates; D002846:Chromatography, Affinity / Q000379:methods; D004355:Drug Stability; D005782:Gels; D011621:Pteridines; D011817:Rabbits; D014365:Tryptophan Hydroxylase / Q000302:isolation & purification* / Q000378:metabolism
D002130:Calcium Phosphates; D005782:Gels; D011621:Pteridines; C005063:2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine; D014365:Tryptophan Hydroxylase