PMID: 6981058May 14, 1982Paper

Specificities and properties of three reduced pyridine nucleotide-flavin mononucleotide reductases coupling to bacterial luciferase

Molecular and Cellular Biochemistry
H Watanabe, J W Hastings

Abstract

Three different NAD(P)H-FMN reductases were extracted from Beneckea harveyi MB-20 cells and separated by DEAE-Sephadex A50 column chromatography. Further purification was achieved by affinity chromatography. In determinations of Km values for NADH, NADPH, and FMN, these three reductases exhibited different specificities and kinetic parameters. One reductase utilizes NADH, whereas a second one utilizes NADPH as the preferred substrate. The third, a newly described reductase species, exhibits about the same reaction rates with NADH and NADPH. The reaction mechanisms of the three enzyme forms have been deduced by steady state kinetic analysis. The highly pure (based on gel electrophoresis) NADPH-FMN reductase still exhibited a low (approximately 2%) activity for NADH, which activity was increased upon storage at 4 degrees but suppressed completely by the replacement of the phosphate buffer with sodium citrate buffer. This high specificity of NADPH-FMN reductase for NADPH under these conditions is useful for the assay of NADPH, notably in systems coupled to bacterial luciferase.

Citations

Jan 5, 2002·Antioxidants & Redox Signaling·S C Tu
Oct 1, 1995·Photochemistry and Photobiology·S C Tu, H I Mager
Dec 29, 2011·Frontiers in Bioscience (Landmark Edition)·Ranjan K Singh, John J Tanner
Jul 15, 1987·Biochemical and Biophysical Research Communications·P MacherouxJ W Hastings
Mar 1, 1987·Applied Biochemistry and Biotechnology·H K Chenault, G M Whitesides
Feb 12, 2008·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Shiao-Chun Tu
Dec 22, 1995·The Journal of Biological Chemistry·F FieschiM Fontecave
Jul 1, 1990·Journal of Bioluminescence and Chemiluminescence·J T LaviT H Stahlberg
Mar 18, 2000·The Journal of Nutrition·S C TuC Jeffers
Apr 28, 1994·Biochimica Et Biophysica Acta·Y IzumotoK Yamamoto

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