Mutagenesis of nitrite reductase from Pseudomonas aeruginosa: tyrosine-10 in the c heme domain is not involved in catalysis

FEBS Letters
F CutruzzolaM Brunori

Abstract

In Pseudomonas aeruginosa, conversion of nitrite to NO in dissimilatory denitrification is catalyzed by the enzyme nitrite reductase (NiR), a homodimer containing a covalently bound c heme and a d1 heme per subunit. We report the purification and characterization of the first single mutant of P. aeruginosa cd1 NiR in which Tyr10 has been replaced by Phe; this amino acid was chosen as a possibly important residue in the catalytic mechanism of this enzyme based on the proposal (Fulop, V., Moir, J.W.B., Ferguson, S.J. and Hajdu, J. (1995) Cell 81, 369-377) that the topologically homologous Tyr25 plays a crucial role in controlling the activity of the cd1 NiR from Thiosphaera pantotropha. Our results show that in P. aeruginosa NiR substitution of Tyr10 with Phe has no effect on the activity, optical spectroscopy and electron transfer kinetics of the enzyme, indicating that distal coordination of the Fe3+ of the d1 heme is provided by different side-chains in different species.

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Citations

May 13, 1999·Biochimica Et Biophysica Acta·F Cutruzzolà
Jan 3, 2003·Protein Expression and Purification·Marzia AreseFrancesca Cutruzzolà
Apr 3, 2001·Current Opinion in Chemical Biology·I Moura, J J Moura
Jul 17, 1998·Current Opinion in Chemical Biology·S J Ferguson
Feb 28, 2001·Proceedings of the National Academy of Sciences of the United States of America·F CutruzzolaM Brunori
Feb 7, 2002·Biochemical and Biophysical Research Communications·Wenliang SunFrancesca Cutruzzolà
May 13, 1999·Biochimica Et Biophysica Acta·N J WatmoughS Spiro
Apr 4, 2014·Chemical Reviews·Luisa B Maia, José J G Moura
Aug 12, 2015·Journal of Bacteriology·Francesca Cutruzzola, Nicole Frankenberg-Dinkel
May 18, 1999·The Journal of Biological Chemistry·D NurizzoM Tegoni
Jun 9, 2020·Acta Crystallographica. Section F, Structural Biology Communications·Thomas Klünemann, Wulf Blankenfeldt
Dec 31, 1997·Microbiology and Molecular Biology Reviews : MMBR·W G Zumft
Feb 12, 2011·Journal of the American Chemical Society·Marina RadoulDaniella Goldfarb

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