Purification and characterization of 6-pyruvoyl tetrahydropterin synthase from human pituitary gland.
6-Pyruvoyl tetrahydropterin synthase, the enzyme that catalyses the conversion of 7,8-dihydroneopterin triphosphate to 6-pyruvoyl tetrahydropterin, was purified 3,330-fold from human pituitary gland with an overall recovery of 30%. The native enzyme has a molecular mass of 68 kD and consists of four identical subunits of 16.5 kD. The pH optimum of the enzyme in Tris/HCl buffer is 7.5. The enzyme is dependent on Mg2+ and NADPH and has a Michaelis-Menten constant of 10 microM for its natural substrate, 7,8-dihydroneopterin triphosphate. The isoelectric point of the human enzyme is 4.3-4.6. The human pituitary gland enzyme is heat instable in contrast to the enzymes from human, rat and salmon liver, and Drosophila head. The amino acid composition showed remarkably high content of acidic amino acids Asp and Glu. The N-terminus was found to be blocked.
Guzman
J
J
Department of Pediatrics, University of Zürich, Switzerland.
Redweik
U
U
Schoedon
G
G
Hunziker
P
P
Wiestler
O D
OD
Heizmann
C W
CW
Blau
N
N
D000429:Alcohol Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D001708:Biopterins / Q000031:analogs & derivatives / Q000096:biosynthesis; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004330:Drosophila / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007150:Immunohistochemistry; D007525:Isoelectric Focusing; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D046911:Macromolecular Substances; D008970:Molecular Weight; D019761:Phosphorus-Oxygen Lyases; D010902:Pituitary Gland / Q000201:enzymology*; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000201:enzymology; D010904:Pituitary Gland, Posterior / Q000166:cytology / Q000201:enzymology; D051381:Rats; D012474:Salmon
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D046911:Macromolecular Substances; D001708:Biopterins; D000429:Alcohol Oxidoreductases; D019761:Phosphorus-Oxygen Lyases; C043497:6-pyruvoyltetrahydropterin synthase; C003402:sapropterin