PMID- 10625700
OWN - NLM
STAT- MEDLINE
DCOM- 20000218
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 275
IP  - 2
DP  - 2000 Jan 14
TI  - Cloning and characterization of a novel human dual flavin reductase.
PG  - 1471-8
AB  - Flavoprotein reductases play a key role in electron transfer in many
      physiological processes. We have isolated a cDNA with strong sequence
      similarities to cytochrome P-450 reductase and nitric-oxide synthase. The cDNA
      encodes a protein of 597 amino acid residues with a predicted molecular mass of
      67 kDa. Northern blot analysis identified a predicted transcript of 3.0 kilobase 
      pairs as well as a larger transcript at 6.0 kilobase pairs, and the gene was
      mapped to chromosome 9q34.3 by fluorescence in situ hybridization analysis. The
      amino acid sequence of the protein contained distinct FMN-, FAD-, and
      NADPH-binding domains, and in order to establish whether the protein contained
      these cofactors, the coding sequence was expressed in insect cells and purified. 
      Recombinant protein bound FMN, FAD, and NADPH cofactors and exhibited a
      UV-visible spectrum with absorbance maxima at 380, 460, and 626 nm. The purified 
      enzyme reduced cytochrome c, with apparent K(m) and k(cat) values of 21 microM
      and 1.3 s(-1), respectively, and metabolized the one-electron acceptors
      doxorubicin, menadione, and potassium ferricyanide. Immunoblot analysis of
      fractionated MCF7 cells with antibodies to recombinant NR1 showed that the enzyme
      is cytoplasmic and highly expressed in a panel of human cancer cell lines, thus
      indicating that this novel reductase may play a role in the metabolic activation 
      of bioreductive anticancer drugs and other chemicals activated by one-electron
      reduction.
FAU - Paine, M J
AU  - Paine MJ
AD  - Imperial Cancer Research Fund Molecular Pharmacology Unit, Biomedical Research
      Centre, University of Dundee, Ninewells Hospital and Medical School, Dundee DD1
      9SY, United Kingdom.
FAU - Garner, A P
AU  - Garner AP
FAU - Powell, D
AU  - Powell D
FAU - Sibbald, J
AU  - Sibbald J
FAU - Sales, M
AU  - Sales M
FAU - Pratt, N
AU  - Pratt N
FAU - Smith, T
AU  - Smith T
FAU - Tew, D G
AU  - Tew DG
FAU - Wolf, C R
AU  - Wolf CR
LA  - eng
SI  - GENBANK/AF199509
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Recombinant Proteins)
RN  - 146-14-5 (Flavin-Adenine Dinucleotide)
RN  - 53-59-8 (NADP)
RN  - 7N464URE7E (Flavin Mononucleotide)
RN  - 9035-42-1 (Cytochromes c1)
RN  - EC 1.5.1.38 (FMN Reductase)
RN  - EC 1.6.- (NADH, NADPH Oxidoreductases)
SB  - IM
MH  - 3T3 Cells
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Binding Sites
MH  - Breast Neoplasms
MH  - Chromosome Mapping
MH  - *Chromosomes, Human, Pair 6
MH  - Cloning, Molecular
MH  - Cytochromes c1/metabolism
MH  - FMN Reductase
MH  - Female
MH  - Flavin Mononucleotide/metabolism
MH  - Flavin-Adenine Dinucleotide/metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Kinetics
MH  - Mice
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - NADH, NADPH Oxidoreductases/chemistry/*genetics/*metabolism
MH  - NADP/metabolism
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Spectrophotometry
MH  - Tumor Cells, Cultured
EDAT- 2000/01/08 09:00
MHDA- 2000/02/26 09:00
CRDT- 2000/01/08 09:00
PHST- 2000/01/08 09:00 [pubmed]
PHST- 2000/02/26 09:00 [medline]
PHST- 2000/01/08 09:00 [entrez]
AID - 10.1074/jbc.275.2.1471 [doi]
PST - ppublish
SO  - J Biol Chem. 2000 Jan 14;275(2):1471-8. doi: 10.1074/jbc.275.2.1471.