The MCD and EPR of the Heme Centers of Nitric Oxide Reductase fromPseudomonas stutzeri:  Evidence That the Enzyme Is Structurally Related to the Heme-Copper Oxidases†

Biochemistry ◽  
1998 ◽  
Vol 37 (11) ◽  
pp. 3994-4000 ◽  
Author(s):  
Myles R. Cheesman ◽  
Walter G. Zumft ◽  
Andrew J. Thomson
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Reductase ◽  
Heme Copper Oxidases

A Low-Redox Potential Heme in the Dinuclear Center of Bacterial Nitric Oxide Reductase: Implications for the Evolution of Energy-Conserving Heme−Copper Oxidases†

Biochemistry ◽  
1999 ◽  
Vol 38 (42) ◽  
pp. 13780-13786 ◽  
Author(s):  
Karin L. C. Grönberg ◽  
M. Dolores Roldán ◽  
Louise Prior ◽  
Gareth Butland ◽  
Myles R. Cheesman ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Redox Potential ◽  
Nitric Oxide Reductase ◽  
Energy Conserving ◽  
Heme Copper Oxidases

scholarly journals Two Conserved Glutamates in the Bacterial Nitric Oxide Reductase Are Essential for Activity but Not Assembly of the Enzyme

2001 ◽  
Vol 183 (1) ◽  
pp. 189-199 ◽  
Author(s):  
Gareth Butland ◽  
Stephen Spiro ◽  
Nicholas J. Watmough ◽  
David J. Richardson
Keyword(s):  
Nitric Oxide ◽  
Paracoccus Denitrificans ◽  
Transmembrane Helix ◽  
Spectroscopic Characterization ◽  
Nitric Oxide Reductase ◽  
Intact Cells ◽  
Heterologous System ◽  
Proton Uptake ◽  
Heme Copper Oxidases

ABSTRACT The bacterial nitric oxide reductase (NOR) is a divergent member of the family of respiratory heme-copper oxidases. It differs from other family members in that it contains an FeB–heme-Fe dinuclear catalytic center rather than a CuB–heme-Fe center and in that it does not pump protons. Several glutamate residues are conserved in NORs but are absent in other heme-copper oxidases. To facilitate mutagenesis-based studies of these residues inParacoccus denitrificans NOR, we developed two expression systems that enable inactive or poorly active NOR to be expressed, characterized in vivo, and purified. These are (i) a homologous system utilizing the cycA promoter to drive aerobic expression of NOR in P. denitrificans and (ii) a heterologous system which provides the first example of the expression of an integral-membrane cytochrome bc complex inEscherichia coli. Alanine substitutions for three of the conserved glutamate residues (E125, E198, and E202) were introduced into NOR, and the proteins were expressed in P. denitrificans and E. coli. Characterization in intact cells and membranes has demonstrated that two of the glutamates are essential for normal levels of NOR activity: E125, which is predicted to be on the periplasmic surface close to helix IV, and E198, which is predicted to lie in the middle of transmembrane helix VI. The subsequent purification and spectroscopic characterization of these enzymes established that they are stable and have a wild-type cofactor composition. Possible roles for these glutamates in proton uptake and the chemistry of NO reduction at the active site are discussed.

scholarly journals Nitric oxide reductase. Purification from Paracoccus denitrificans with use of a single column and some characteristics

1991 ◽  
Vol 266 (17) ◽  
pp. 10899-10905
Author(s):  
M. Dermastia ◽  
T. Turk ◽  
T.C. Hollocher
Keyword(s):  
Nitric Oxide ◽  
Paracoccus Denitrificans ◽  
Nitric Oxide Reductase ◽  
Single Column

scholarly journals Kinetics of electron transfer from NADH to the Escherichia coli nitric oxide reductase flavorubredoxin

FEBS Journal ◽  
2006 ◽  
Vol 274 (3) ◽  
pp. 677-686 ◽  
Author(s):  
João B. Vicente ◽  
Francesca M. Scandurra ◽  
João V. Rodrigues ◽  
Maurizio Brunori ◽  
Paolo Sarti ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Escherichia Coli ◽  
Electron Transfer ◽  
Nitric Oxide Reductase ◽  
Kinetics Of

The co-immobilization of P450-type nitric oxide reductase and glucose dehydrogenase for the continuous reduction of nitric oxide via cofactor recycling

2016 ◽  
Vol 85 ◽  
pp. 71-81 ◽  
Author(s):  
Seike Garny ◽  
Natasha Beeton-Kempen ◽  
Isak Gerber ◽  
Jan Verschoor ◽  
Justin Jordaan
Keyword(s):  
Nitric Oxide ◽  
Glucose Dehydrogenase ◽  
Nitric Oxide Reductase ◽  
Cofactor Recycling

scholarly journals 2P110 Elucidation of NO reduction reaction using Zn-substituted bacterial nitric oxide reductase(The 48th Annual Meeting of the Biophysical Society of Japan)

2010 ◽  
Vol 50 (supplement2) ◽  
pp. S101
Author(s):  
Norihiro Okada ◽  
Takehiko Tosha ◽  
Masaki Horitani ◽  
Yoshitsugu Shiro
Keyword(s):  
Nitric Oxide ◽  
Annual Meeting ◽  
No Reduction ◽  
Reduction Reaction ◽  
Nitric Oxide Reductase ◽  
Biophysical Society

Reaction of Carbon Monoxide with the Reduced Active Site of Bacterial Nitric Oxide Reductase†

Biochemistry ◽  
2001 ◽  
Vol 40 (44) ◽  
pp. 13361-13369 ◽  
Author(s):  
Janneke H. M. Hendriks ◽  
Louise Prior ◽  
Adam R. Baker ◽  
Andrew J. Thomson ◽  
Matti Saraste ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Carbon Monoxide ◽  
Active Site ◽  
Nitric Oxide Reductase

scholarly journals 2P080 Reaction Mechanism of Nitric Oxide Reductase Cytochrome P450nor from Fusarium oxysporum

2004 ◽  
Vol 44 (supplement) ◽  
pp. S129
Author(s):  
K. Tsukamoto ◽  
T. Watanabe ◽  
U. Nagashima ◽  
Y. Akiyama
Keyword(s):  
Nitric Oxide ◽  
Reaction Mechanism ◽  
Fusarium Oxysporum ◽  
Nitric Oxide Reductase

scholarly journals A new assay for nitric oxide reductase reveals two conserved glutamate residues form the entrance to a proton-conducting channel in the bacterial enzyme

2006 ◽  
Vol 401 (1) ◽  
pp. 111-119 ◽  
Author(s):  
Faye H. Thorndycroft ◽  
Gareth Butland ◽  
David J. Richardson ◽  
Nicholas J. Watmough
Keyword(s):  
Nitric Oxide ◽  
Paracoccus Denitrificans ◽  
Nitric Oxide Reductase ◽  
Transmembrane Helices ◽  
Level Of Activity ◽  
Bacterial Enzyme ◽  
Cytoplasmic Membranes ◽  
Membrane Bound ◽  
Proton Movement ◽  
High Level

A specific amperometric assay was developed for the membrane-bound NOR [NO (nitric oxide) reductase] from the model denitrifying bacterium Paracoccus denitrificans using its natural electron donor, pseudoazurin, as a co-substrate. The method allows the rapid and specific assay of NO reduction catalysed by recombinant NOR expressed in the cytoplasmic membranes of Escherichia coli. The effect on enzyme activity of substituting alanine, aspartate or glutamine for two highly conserved glutamate residues, which lie in a periplasmic facing loop between transmembrane helices III and IV in the catalytic subunit of NOR, was determined using this method. Three of the substitutions (E122A, E125A and E125D) lead to an almost complete loss of NOR activity. Some activity is retained when either Glu122 or Glu125 is substituted with a glutamine residue, but only replacement of Glu122 with an aspartate residue retains a high level of activity. These results are interpreted in terms of these residues forming the mouth of a channel that conducts substrate protons to the active site of NOR during turnover. This channel is also likely to be that responsible in the coupling of proton movement to electron transfer during the oxidation of fully reduced NOR with oxygen [U. Flock, N. J. Watmough and P. Ädelroth (2005) Biochemistry 44, 10711–10719].

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