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4 Pro-R Hydrogen of NADPH was Abstracted for Enzymatic Hydride Transfer by N-Ethylmaleimide Reductase of Yarrowia lipolytica
- Source :
- The Tohoku Journal of Experimental Medicine. 181:447-457
- Publication Year :
- 1997
- Publisher :
- Tohoku University Medical Press, 1997.
-
Abstract
- We studied the steric course of the reaction catalyzed by the N-ethylmaleimide (NEM) reductase of Yarrowia (Candida) lipolytica (Y. lipolytica), using 4R-[4-2H1]NADPH and 4S-[4-2H1]NADPH as cofactors and N-ethylcitraconimide as substrate. Active substrates and inhibitors of NEM reductase and its subcellular distribution were also investigated to clarify the biochemical properties of this enzyme. NEM reductase catalyzes the reduction of N-ethylmaleimide to N-ethylsuccinimide with NAD(P)H as the cofactor. Several maleimide and cyclopentenone derivatives tested were also active substrates for NEM reductase of Y. lipolytica. Some pyrazolone derivatives, particularly 1-phenyl-5-pyrazolone, were found to be effective inhibitors of NEM reductase. Subcellular localization of NEM reductase was carried out using protoplast formation and differential centrifugation. Ninety-eight percent of the NEM reductase activity was recovered in the cytosolic fraction, indicating that NEM reductase in Y. lipolytica was the cytosolic enzyme. We also determined the stereochemical specificity of the reduction of N-ethylcitraconimide by NEM reductase in Y. lipolytica, showing that 4 Pro-R hydrogen of NADPH was abstracted for enzymatic hydride transfer by NEM reductase, and two hydrogen atoms from NADPH and H2O added to opposite faces of the double bond of N-ethylcitraconimide.
- Subjects :
- Stereochemistry
Stereoisomerism
Cyclopentanes
Reductase
General Biochemistry, Genetics and Molecular Biology
Cofactor
Substrate Specificity
Maleimides
chemistry.chemical_compound
heterocyclic compounds
Enzyme Inhibitors
Candida
chemistry.chemical_classification
biology
N-Ethylmaleimide
Substrate (chemistry)
Yarrowia
General Medicine
Deuterium
biology.organism_classification
Enzyme
chemistry
Biochemistry
Ethylmaleimide
biology.protein
Pyrazoles
NAD+ kinase
Oxidoreductases
Oxidation-Reduction
NADP
Hydrogen
Subjects
Details
- ISSN :
- 13493329 and 00408727
- Volume :
- 181
- Database :
- OpenAIRE
- Journal :
- The Tohoku Journal of Experimental Medicine
- Accession number :
- edsair.doi.dedup.....4ac3244d074c2b108b42cc8f4224d87d