PROPERTIES OF TYROSINE HYDROXYLASE IN PERIPHERAL NERVES

Abstract

Abstract—Approximately 80 per cent of tyrosine hydroxylase activity in bovine mandibular nerve and rabbit sciatic nerve was soluble, and the rest of the activity was particle‐bound. The soluble enzyme in bovine mandibular nerve was isolated by ammonium sulphate fractionation (25–35 per cent saturation). The enzyme had a pH optimum at 5·9 in Tris‐acetate buffer, and at 6·5 in Tris‐HCl or phosphate buffer. The enzyme required a tetrahydropteridine cofactor. Km values toward various tetrahydropteridines such as l‐erythro‐tetrahydrobiopterin (a probable natural cofactor), 2‐amino‐4‐hydroxy‐6‐methyltetrahydropteridine, and 2‐amino‐4‐hydroxy‐6,7‐dimethyltetrahydropteridine were 2 × 10−5m, 5 × 10−5m and 4 × 10−4m, respectively. The Km value for tyrosine at 1 × 10−3m‐2‐amino‐4‐hydroxy‐6‐methyltetrahydropteridine as a cofactor was 5 × 10−5m. The enzyme activity was markedly stimulated with Fe2+ or catalase, but Fe2+ gave higher activity. The activity was inhibited with α, α′‐dipyridyl, l‐α‐methyl‐p‐tyrosine, and various catecholamines. Among catecholamines, dopamine was the most potent inhibitor. l‐5‐Hydroxytryptophan was an inhibitor as potent as dopamine. Neither d‐5‐hydroxytryptophan nor 5‐hydroxytryptamine inhibited the enzyme. The inhibition by l‐5‐hydroxytryptophan was partially competitive with tetrahydrobiopterin at concentrations higher than 9 × 10−5m, and partially uncompetitive at concentrations lower than 9 × 10−5m. The addition of heparin or lysolecithin did not affect enzyme activity with tetrahydrobiopterin as cofactor.