RECOMBINANT HUMAN TYROSINE-HYDROXYLASE TYPES-1-4 SHOW REGULATORY KINETIC-PROPERTIES FOR THE NATURAL (6R)-TETRAHYDROBIOPTERIN COFACTOR

被引:37
作者
NASRIN, S
ICHINOSE, H
HIDAKA, H
NAGATSU, T
机构
[1] FUJITA HLTH UNIV,SCH MED,INST COMPREHENS MED SCI,TOYOAKE,AICHI 47011,JAPAN
[2] NAGOYA UNIV,SCH MED,DEPT PHARMACOL,NAGOYA,AICHI 466,JAPAN
关键词
ESCHERICHIA COLI; KINETIC PROPERTIES; RECOMBINANT HUMAN ENZYMES; (6R)-TETRAHYDROBIOPTERIN; TYROSINE HYDROXYLASE TYPES 1-4;
D O I
10.1093/oxfordjournals.jbchem.a124537
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Human tyrosine hydroxylase (hTH) exists in four isoforms. The four recombinant hTH isoenzymes types 1-4 (hTH1-4) produced in and purified from Escherichia coli showed regulatory kinetic properties for the natural(6R)-L-erythro-tetrahydrobiopterin (RBPH4) as a cofactor. In contrast, the unnatural cofactor (6S)-L-erythro-tetrahydrobiopterin (SBPH4) and a synthetic cofactor (6RS)-methyl-tetrahydropterin (6MPH4) showed usual kinetic characteristics with each of hTH1-4. Substrate inhibition by tyrosine was observed for each of hTH1-4 with natural RBPH4. Two different K-m values for pterin cofactor were observed at a high concentration (200 mu M) of L-tyrosine only with natural RBPH4, in contrast to a single K-m value for unnatural SBPH4 or synthetic 6MPH4. The present results suggest that in the presence of relatively high concentrations (approximately 100 mu M) of tyrosine in vivo, RBPH4 cofactor may have a regulatory role for the activity of all four human isoenzymes in vivo. We also found that recombinant hTH1 and 3 were more unstable than hTH2 and 4, suggesting that the 4-amino-acid insertion in hTH2 and 4 may be responsible for the relative stability of hTH2 and 4 isoenzymes.
引用
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页码:393 / 398
页数:6
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