Enzymes related to catecholamine biosynthesis in Tetrahymena pyriformis. Presence of GTP cyclohydrolase I.
We first identified GTP cyclohydrolase I activity (EC 3.5.4.16) in the ciliated protozoa, Tetrahymena pyriformis. The Vmax value of the enzyme in the cellular extract of T. pyriformis was 255 pmol mg-1 protein h-1. Michaelis-Menten kinetics indicated a positive cooperative binding of GTP to the enzyme. The GTP concentration producing half-maximal velocity was 0.8 mM. By high-performance liquid chromatography (HPLC) with fluorescence detection, a major peak corresponding to D-monapterin (2-amino-4-hydroxy-6-[(1'R,2'R)-1',2',3'-trihydroxypropyl]pteridin e, D-threo-neopterin) and minor peaks of D-erythro-neopterin and L-erythro-biopterin were found to be present in the cellular extract of Tetrahymena. Thus, it is strongly suggested that Tetrahymena converts GTP into unconjugated pteridine derivatives. In this study, dopamine was detected as the major catecholamine, while neither epinephrine nor norepinephrine was identified. Indeed, this protozoa was shown to possess the activity of a dopamine synthesizing enzyme, aromatic L-amino acid decarboxylase. On the other hand, activities of tyrosine hydroxylase or tyrosinase which converts tyrosine into dopa, the substrate of aromatic L-amino acid decarboxylase, could not be detected in this protozoa. Furthermore, neither dopamine beta-hydroxylase activity nor phenylethanolamine N-methyltransferase activity could be identified by the HPLC methods.
Nomura
T
T
Department of Pharmacology, Fujita Health University School of Medicine, Aichi, Japan.
Tazawa
M
M
Ohtsuki
M
M
Sumi-Ichinose
C
C
Hagino
Y
Y
Ota
A
A
Nakashima
A
A
Mori
K
K
Sugimoto
T
T
Ueno
O
O
Nozawa
Y
Y
Ichinose
H
H
Nagatsu
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000378:metabolism; D002395:Catecholamines / Q000096:biosynthesis*; D004298:Dopamine / Q000096:biosynthesis; D006136:GTP Cyclohydrolase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D011621:Pteridines / Q000378:metabolism; D013769:Tetrahymena pyriformis / Q000201:enzymology* / Q000378:metabolism
D002395:Catecholamines; D011621:Pteridines; D006160:Guanosine Triphosphate; D006136:GTP Cyclohydrolase; D001142:Aromatic-L-Amino-Acid Decarboxylases; D004298:Dopamine