Molecular cloning and amino acid sequence of human 5-lipoxygenase.
5-Lipoxygenase (EC 1.13.11.34), a Ca2+-and ATP-requiring enzyme, catalyzes the first two steps in the biosynthesis of the peptidoleukotrienes and the chemotactic factor leukotriene B4. A cDNA clone corresponding to 5-lipoxygenase was isolated from a human lung lambda gt11 expression library by immunoscreening with a polyclonal antibody. Additional clones from a human placenta lambda gt11 cDNA library were obtained by plaque hybridization with the 32P-labeled lung cDNA clone. Sequence data obtained from several overlapping clones indicate that the composite cDNAs contain the complete coding region for the enzyme. From the deduced primary structure, 5-lipoxygenase encodes a 673 amino acid protein with a calculated molecular weight of 77,839. Direct analysis of the native protein and its proteolytic fragments confirmed the deduced composition, the amino-terminal amino acid sequence, and the structure of many internal segments. 5-Lipoxygenase has no apparent sequence homology with leukotriene A4 hydrolase or Ca2+ -binding proteins. RNA blot analysis indicated substantial amounts of an mRNA species of approximately equal to 2700 nucleotides in leukocytes, lung, and placenta.
Matsumoto
T
T
Department of Physiological Chemistry, Karolinska Institutet, Stockholm, Sweden.
Funk
C D
CD
Rådmark
O
O
Höög
J O
JO
Jörnvall
H
H
Samuelsson
B
B
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001094:Arachidonate 5-Lipoxygenase / Q000235:genetics*; D001091:Arachidonate Lipoxygenases / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes; D005260:Female; D005796:Genes; D006801:Humans; D008168:Lung / Q000201:enzymology; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D010920:Placenta / Q000201:enzymology; D012333:RNA, Messenger / Q000235:genetics
D000596:Amino Acids; D012333:RNA, Messenger; D004247:DNA; D001091:Arachidonate Lipoxygenases; D001094:Arachidonate 5-Lipoxygenase; D004262:DNA Restriction Enzymes