Reduction of lipid peroxidation products and advanced glycation end-product precursors by cyanobacterial aldo-keto reductase AKR3G1—a founding member of the AKR3G subfamily.
The purpose of this study was to investigate the origin and function of the aldo-keto reductase (AKR) superfamily as enzymes involved in the detoxification of xenobiotics. We used the cyanobacterium Synechocystis sp. PCC 6803 as a model organism and sequence alignments to find bacterial AKRs with highest identity to human enzymes. Disappearance of NADPH was monitored spectrophotometrically to calculate enzymatic activity. The molecular weight of the native protein was determined by size exclusion chromatography. Substrate docking was performed by SwissDock. Sequence alignments identified the NADPH-dependent AKR3G1 having 41.5 and 40% identity with the human enzymes AKR1B1 and AKR1B10, respectively. Highest enzymatic efficiency was observed with 4-oxonon-2-enal (4-ONE; k(cat)/K(m), 561 s(-1) mM(-1)) and 4-hydroxynonenal (k(cat)/K(m), 26.5 s(-1) mM(-1)), respectively. P74308 is the most efficient enzyme for 4-ONE discovered until now. Cooperativity of this monomeric enzyme was observed with some substrates. Enzyme inactivation or oligomerization as possible explanations for nonhyperbolic enzyme kinetics were ruled out by Selwyn's test and gel filtration. The role of the little investigated carbonyl-reducing enzymes in detoxification seems to be in fact a very old process with rarely observed nonhyperbolic enzyme kinetics as an adaptation mechanism to higher concentrations of reactive oxygen species.
Institute of Toxicology and Pharmacology for Natural Scientists, University Medical School Schleswig-Holstein, Campus Kiel, Kiel, Germany hintzpeter@toxi.uni-kiel.de.
Jan
J
Hintzpeter
Institute of Toxicology and Pharmacology for Natural Scientists, University Medical School Schleswig-Holstein, Campus Kiel, Kiel, Germany.
Hans-Joerg
HJ
Martin
Institute of Toxicology and Pharmacology for Natural Scientists, University Medical School Schleswig-Holstein, Campus Kiel, Kiel, Germany.
Edmund
E
Maser
D000449:Aldehyde Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism; D000074408:Aldo-Keto Reductases; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain; D017127:Glycation End Products, Advanced / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008024:Ligands; D015227:Lipid Peroxidation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D046939:Synechocystis / Q000201:enzymology* / Q000235:genetics
C403894:4-oxo-2-nonenal; D000447:Aldehydes; D001426:Bacterial Proteins; D017127:Glycation End Products, Advanced; D008024:Ligands; D011994:Recombinant Proteins; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase; C027576:4-hydroxy-2-nonenal
carbonyl reduction; detoxification; enzyme kinetics