Expression of human leukotriene A4 hydrolase cDNA in Escherichia coli
Open Access
- 14 March 1988
- journal article
- Published by Wiley in FEBS Letters
- Vol. 229 (2) , 279-282
- https://doi.org/10.1016/0014-5793(88)81140-2
Abstract
The cDNA clone encoding human leukotriene A4 hydrolase was inserted into a vector pUC9 and expressed in Escherichia coli as a fusion protein containing the first 10 amino acid residues derived from a vector. The leukotriene A4 hydrolase activity was recovered in the soluble fraction of the transformants. The purified enzyme showed kinetic properties similar to the native enzyme, including inactivation by the substrate and sulfhydryl‐modifying reagents. The results demonstrate that a protein with an M r of 70000 was expressed in Escherichia coli with a full enzyme activity and structural fidelity. Acquisition of the expression system makes it feasible to elucidate the reaction mechanism of the enzyme.Keywords
This publication has 15 references indexed in Scilit:
- Studies on transformation of Escherichia coli with plasmidsPublished by Elsevier ,2006
- Molecular cloning of a cDNA coding for human leukotriene A4 hydrolase. Complete primary structure of an enzyme involved in eicosanoid synthesis.Journal of Biological Chemistry, 1987
- Molecular cloning and amino acid sequence of leukotriene A4 hydrolase.Proceedings of the National Academy of Sciences, 1987
- Leukotrienes and Lipoxins: Structures, Biosynthesis, and Biological EffectsScience, 1987
- Characterization of leukotriene A4 synthase from murine mast cells: evidence for its identity to arachidonate 5-lipoxygenase.Proceedings of the National Academy of Sciences, 1986
- ARACHIDONIC ACID METABOLISMAnnual Review of Biochemistry, 1986
- Single protein from human leukocytes possesses 5-lipoxygenase and leukotriene A4 synthase activities.Proceedings of the National Academy of Sciences, 1986
- Leukotriene A4 hydrolase in human leukocytes. Purification and properties.Journal of Biological Chemistry, 1984
- Enzyme with dual lipoxygenase activities catalyzes leukotriene A4 synthesis from arachidonic acid.Proceedings of the National Academy of Sciences, 1984
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970