Correlation of the genotoxic activation and kinetic properties of Salmonella enterica serovar Typhimurium nitroreductases SnrA and cnr with the redox potentials of nitroaromatic compounds and quinones.
Bacterial nitroreductases (NRs) catalyse the oxygen-insensitive reduction of several nitro-substituted compounds and quinones. SnrA and cnr NRs have been previously identified in Salmonella enterica serovar Typhimurium; they reduce several environmental nitro compounds that display mutagenic activity in the Ames test. Although some of their biochemical properties have been reported, the substrate specificity of each protein over mutagenic nitro compounds is unknown; even more, the possible relationship between their capacity to activate nitro compounds into mutagens and the redox properties of putative substrates has been poorly investigated. We have purified SnrA and cnr and investigated their capacity to activate several mutagens in the Ames test as well as their kinetic parameters K(m) and V(max). Our results show that SnrA and cnr are able to activate 2,7-dinitrofluorene with the same efficiency and a similar mutagenic potency in the YG7132 tester strain; 1-nitropyrene and 1,3-dinitropyrene were efficiently activated by cnr, whereas 1,8-dinitropyrene, 1,6-dinitropyrene and 2-nitrofluorene were scarcely activated by either NR. The mutagenic potency of nitro compounds obtained in the presence of either enzyme correlates with their redox potential reported in the literature. On the other hand, a good correlation was obtained between the catalytic efficiency (V(max)/K(m)) of the purified cnr with the redox potential of eight molecules including nitro-substituted compounds and quinones. No correlation between redox potential and catalytic efficiency by SnrA was observed, suggesting that factors other than redox potential such as the structure of the compounds are involved in the catalytic efficiency of SnrA.
Salamanca-Pinzón
S G
SG
Departamento de Medicina Genómica y Toxicología Ambiental, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Apartado Postal 70228, México, D.F. 04510, México.
Camacho-Carranza
R
R
Hernández-Ojeda
S L
SL
Frontana-Uribe
B A
BA
Espitia-Pinzón
C I
CI
Espinosa-Aguirre
J J
JJ
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D055162:Biocatalysis / Q000187:drug effects; D055664:Electrochemical Techniques; D004789:Enzyme Activation / Q000187:drug effects; D006841:Hydrocarbons, Aromatic / Q000633:toxicity*; D007700:Kinetics; D009152:Mutagenicity Tests; D009574:Nitro Compounds / Q000633:toxicity*; D009601:Nitroreductases / Q000302:isolation & purification / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D011809:Quinones / Q000633:toxicity*; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000201:enzymology*
D001426:Bacterial Proteins; D006841:Hydrocarbons, Aromatic; D009574:Nitro Compounds; D011809:Quinones; D009601:Nitroreductases