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Expression of human 5-lipoxygenase cDNA in Escherichia coli
Authors:M Noguchi  T Matsumoto  M Nakamura  M Noma
Affiliation:Life Science Research Laboratory, Japan Tobacco Inc., Yokohama.
Abstract:The cis/trans interconversion of Glt-Ala-Ala-Pro-Phe-4-nitroanilide and Glt-Ala-Gly-Pro-Phe-4-nitroanilide was studied both enzymatically and nonenzymatically by measuring kinetic β-deuterium isotope effects. The hydrogen atom at the -carbon atom of the Xaa residue within the Xaa-Pro moiety was substituted by deuterium. In the nonenzymatic case the transition state of rotation is reflected by kH/kD > 1. When catalysed by 17 kDa PPIase the same bond rotation is characterized by kH/kD < 1. This suggests a covalent mechanism of catalysis which involves an approximately tetravalent carbon of the prolyl imidic bond for the transition state of reaction.
Keywords:cis/trans interconversion   Prolyl imidic bond   Deuterium isotope effect   Prolyl cis/trans-isomerase   Cyclophilin   Cyclosporin
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