Identification in various chlorate-resistant mutants of a protein involved in the activation of nitrate reductase in the soluble fraction of a chlA mutant of Escherichia coli K-12.
We report some properties of Protein PA which has been isolated from the soluble fraction of a chlB mutant after anaerobic growth in the presence of KNO3. This protein has been identified by its capacity to reactivate nitrate reductase present in the soluble fraction of a chlA mutant by the complementation process. The presence of active Protein PA in the chlB mutant is independent of the presence of oxygen or of nitrate during growth. In contrast, the addition of sodium tungstate to the growth medium leads to the formation of inactive Protein PA which is not able to activate nitrate reductase in the chlA-soluble extract by complementation. Inactive Protein PA has been quantitated immunologically. The partial purification of Protein PA has been achieved from various chlorate-resistant mutants (chlA-chlG). The establishment of particular complementation systems comprising the soluble extracts of chlA or chlB mutants and partially purified Protein PA from soluble fractions of different chlorate-resistant mutants, has allowed the quantitative estimation of this protein. The analysis by 'rocket immunoelectrophoresis' using an antiserum specific for Protein PA has shown that inactive Protein PA is present in approximately equivalent amounts in the chlA, chlE, chlG and chlD mutants.
D001426:Bacterial Proteins / Q000032:analysis*; D002704:Chlorates / Q000494:pharmacology*; D004352:Drug Resistance, Microbial; D004789:Enzyme Activation; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D008982:Molybdenum / Q000494:pharmacology; D009154:Mutation; D009565:Nitrate Reductases / Q000032:analysis*; D014414:Tungsten / Q000494:pharmacology; D017973:Tungsten Compounds
D001426:Bacterial Proteins; D002704:Chlorates; D017973:Tungsten Compounds; D008982:Molybdenum; C024687:sodium molybdate(VI); D009565:Nitrate Reductases; C021101:tungstic(VI) acid; D014414:Tungsten