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Paracoccus pantotrophus cytochrome cd(1) is a nitrite reductase found in the periplasm of many denitrifying bacteria. It catalyzes the reduction of nitrite to nitric oxide during the denitrification part of the biological nitrogen cycle. Previous studies of early millisecond intermediates in the nitrite reduction reaction have shown, by comparison with pH 7.0, that at the optimum pH, approximately pH 6, the earliest intermediates were lost in the dead time of the instrument. Access to early time points (approximately 100 micros) through use of an ultra-rapid mixing device has identified a spectroscopically novel intermediate, assigned as the Michaelis complex, formed from reaction of fully reduced enzyme with nitrite. Spectroscopic observation of the subsequent transformation of this species has provided data that demand reappraisal of the general belief that the two subunits of the enzyme function independently.

Original publication

DOI

10.1074/jbc.M804493200

Type

Journal article

Journal

J Biol Chem

Publication Date

10/10/2008

Volume

283

Pages

27403 - 27409

Keywords

Cytochromes, Hydrogen-Ion Concentration, Nitrite Reductases, Nitrites, Oxidation-Reduction, Paracoccus, Periplasmic Proteins, Time Factors