The human tyrosine hydroxylase gene.

1. Tyrosine hydroxylase (TH) is a rate-limiting enzyme for catecholamine biosynthesis, and it is a pterin-requiring monooxygenase. Both cDNAs and genomic DNA of human TH have been cloned and the nucleotide sequence has been determined. 2. Four similar but distinct mRNAs encode human TH. The results of Southern blot analysis and the nucleotide sequence of the human TH genomic DAN indicate that the four types of human TH mRNA are produced through alternative splicing from a single gene. 3. The human TH gene was split into 4 exons and 13 introns. The 12-bp insertion sequence is encoded by the 3'-terminal portion of the first exon. The 81-bp insertion sequence corresponds to the second exon. Two kinds of alternative splicing are involved: the alternative use of two donor sites in the first exon and the inclusion/exclusion of the second exon. 4. The four types (type 1-4) were expressed in COS cells, and all had enzymatic activities. The type 1 enzyme had the highest homospecific activity (activity per enzyme protein), the values for the other enzymes ranging from 30 to 40%. The Km values of the four types for L-tyrosine and 6-methyl-5,6,7,8-tetrahydropterin were similar.