Expression of human 5-lipoxygenase cDNA in Escherichia coli.
A cDNA for human 5-lipoxygenase (5LO) was inserted into the vector pKC (constructed from pKK223-3 by replacing its replication origin with that of pUC18) and expressed in Escherichia coli. The enzyme expressed was purified to homogeneity from the cellular soluble fraction. The purified enzyme showed both 5LO and leukotriene A4 synthase activities, which were stimulated by Ca2+ and ATP. Its molecular mass (78 kDa) and NH2-terminal sequence were identical with those of 5LO purified from human leukocytes. The availability of the expression system will facilitate further studies on its regulation and the reaction mechanism of the enzyme.
Noguchi
M
M
Life Science Research Laboratory, Japan Tobacco Inc., Yokohama.
Matsumoto
T
T
Nakamura
M
M
Noma
M
M
D000255:Adenosine Triphosphate / Q000494:pharmacology; D001094:Arachidonate 5-Lipoxygenase / Q000235:genetics*; D001091:Arachidonate Lipoxygenases / Q000235:genetics*; D002118:Calcium / Q000494:pharmacology; D004247:DNA / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D010957:Plasmids
D000255:Adenosine Triphosphate; D004247:DNA; D001091:Arachidonate Lipoxygenases; D001094:Arachidonate 5-Lipoxygenase; D002118:Calcium