Molecular cloning of human platelet thromboxane A synthase.
Complementary DNA coding for thromboxane A synthase was amplified by polymerase chain reaction using primers synthesized according to the partial amino acid sequences of human platelet thromboxane A synthase (Nüsing, R., Schneider-Voss, S., and Ullrich, V. (1990) Arch. Biochem. Biophys. 280, 325-330) and cloned into pBluescript SK II(-). The primary structure of human platelet enzyme was deduced from the nucleotide sequence of the cDNA. The enzyme is composed of 533 amino acids with a molecular weight of 60,487. The primary structure of the enzyme exhibited a 34-36% homology to the amino acid sequences of cytochrome P450s classified in the P450 III gene family. The highly conserved cysteine-containing sequence involved in the heme-binding site of P450 was found near the carboxyl terminus (residues 472-492). The size of the major thromboxane A synthase mRNA from human platelets and human erythroleukemia cells was estimated to be approximately 2.2 kilobases by RNA blot analysis.
Yokoyama
C
C
Department of Pharmacology, National Cardiovascular Center Research Institute, Osaka, Japan.
Miyata
A
A
Ihara
H
H
Ullrich
V
V
Tanabe
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D001792:Blood Platelets / Q000201:enzymology*; D002460:Cell Line; D003001:Cloning, Molecular; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011061:Poly A / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger; D015183:Restriction Mapping; D013930:Thromboxane-A Synthase / Q000235:genetics*
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D013930:Thromboxane-A Synthase