Investigation of the structure of oxidized Pseudomonas aeruginosa cytochrome c-551 by NMR: Comparison of observed paramagnetic shifts and calculated pseudocontact shifts

Biochemistry ◽  
1993 ◽  
Vol 32 (43) ◽  
pp. 11516-11523 ◽  
Author(s):  
Russell Timkovich ◽  
Mengli Cai
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cytochrome C ◽  
Pseudocontact Shifts ◽  
Paramagnetic Shifts

scholarly journals The amino acid sequence of cytochromes c-551 from three species of Pseudomonas

1973 ◽  
Vol 131 (3) ◽  
pp. 485-498 ◽  
Author(s):  
R. P. Ambler ◽  
Margaret Wynn
Keyword(s):  
Amino Acids ◽  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Cytochrome C ◽  
Amino Acid Sequence ◽  
Amino Acid Sequences ◽  
Mitochondrial Cytochrome ◽  
Cytochromes C ◽  
Lending Library ◽  
Single Peptide

The amino acid sequences of the cytochromes c-551 from three species of Pseudomonas have been determined. Each resembles the protein from Pseudomonas strain P6009 (now known to be Pseudomonas aeruginosa, not Pseudomonas fluorescens) in containing 82 amino acids in a single peptide chain, with a haem group covalently attached to cysteine residues 12 and 15. In all four sequences 43 residues are identical. Although by bacteriological criteria the organisms are closely related, the differences between pairs of sequences range from 22% to 39%. These values should be compared with the differences in the sequence of mitochondrial cytochrome c between mammals and amphibians (about 18%) or between mammals and insects (about 33%). Detailed evidence for the amino acid sequences of the proteins has been deposited as Supplementary Publication SUP 50015 at the National Lending Library for Science and Technology, Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1973), 131, 5.

scholarly journals A reinvestigation of the covalent structure of Pseudomonas aeruginosa cytochrome c peroxidase

FEBS Letters ◽  
1995 ◽  
Vol 377 (2) ◽  
pp. 145-149 ◽  
Author(s):  
Bart Samyn ◽  
Kathleen Van Craenenbroeck ◽  
Lina De Smet ◽  
Isabel Vandenberghe ◽  
Graham Pettigrew ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cytochrome C ◽  
Cytochrome C Peroxidase ◽  
Covalent Structure

Comparison of reduced and oxidized yeast iso-1-cytochrome c using proton paramagnetic shifts

Biochemistry ◽  
1991 ◽  
Vol 30 (7) ◽  
pp. 1928-1934 ◽  
Author(s):  
Yuan Gao ◽  
Jonathan Boyd ◽  
Gary J. Pielak ◽  
Robert J. P. Williams
Keyword(s):  
Cytochrome C ◽  
Paramagnetic Shifts

Nitrite activates the transcription of the Pseudomonas aeruginosa nitrite reductase and cytochrome c -551 operon under anaerobic conditions

FEBS Letters ◽  
1991 ◽  
Vol 288 (1-2) ◽  
pp. 227-228 ◽  
Author(s):  
Hiroyuki Arai ◽  
Yasuo Igarashi ◽  
Tohru Kodama
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cytochrome C ◽  
Nitrite Reductase ◽  
Anaerobic Conditions

pH and Temperature Effect on the Absorption Spectra of Pseudomonas aeruginosa Cytochrome c-551 Solution

1994 ◽  
Vol 98 (17) ◽  
pp. 4726-4728 ◽  
Author(s):  
Yongfang Li ◽  
Kenichi Imaeda ◽  
Hiroo Inokuchi
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cytochrome C ◽  
Temperature Effect ◽  
Absorption Spectra

scholarly journals The nature of species prepared by photolysis of half-reduced, fully reduced and fully reduced carbonmonoxy-cytochrome c-551 peroxidase from Pseudomonas aeruginosa

1984 ◽  
Vol 223 (2) ◽  
pp. 379-391 ◽  
Author(s):  
C Greenwood ◽  
N Foote ◽  
J Peterson ◽  
A J Thomson
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Fine Structure ◽  
Low Temperature ◽  
Cytochrome C ◽  
Activation Barrier ◽  
Magnetic Circular Dichroism ◽  
High Potential ◽  
Quantum Mechanical ◽  
Methionine Residue ◽  
Magnetic Circular Dichroism Spectroscopy

The half-reduced, fully reduced and fully reduced CO-bound forms of the enzyme cytochrome c-551 peroxidase isolated from Pseudomonas aeruginosa were examined by a combination of low-temperature absorption and magnetic-circular-dichroism spectroscopy. Deliberate low-temperature (4.2K) photolysis of these forms of the enzyme, in all of which the high-potential haem is in the ferrous state, revealed that this haem group, assigned to have a histidine-methionine ligand set, is photosensitive. The photolabile ligand is most likely to be the methionine residue, and the product of photolysis, namely the high-spin (S = 2) ferrous form, is stable at low temperature (4.2K). Warming to approx. 20K allows thermal recombination to occur, restoring the low-spin (S = 0) state. The low-potential haem (bis-histidine ligation) is photoinert in both ferric and ferrous states; however, the photosensitive CO adduct of this centre cannot be maintained as the photolysed (S = 2) product at 4.2K. This surprising observation may be due to quantum-mechanical tunnelling of the CO through the activation barrier even at 4.2K, implying that the activation barrier to thermal recombination is both narrow and low. Low-temperature absorption spectroscopy reveals that the high-potential haem has a very characteristic low-spin ferrous spectrum with intense highly structured beta- and split alpha-bands, whereas the spectrum of the low-potential ferrous haem contains alpha- and beta-bands devoid of fine structure.

Activation of the cytochrome c peroxidase of Pseudomonas aeruginosa. The role of a heme-linked protein loop: A mutagenesis studies

2007 ◽  
Vol 101 (8) ◽  
pp. 1133-1139 ◽  
Author(s):  
Hsi-Chen Hsiao ◽  
Svetlana Boycheva ◽  
Nicholas J. Watmough ◽  
Thomas Brittain
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cytochrome C ◽  
Cytochrome C Peroxidase

scholarly journals Cytochrome c-551 and azurin oxidation catalysed by Pseudomonas aeruginosa cytochrome oxidase. A steady-state kinetic study

1985 ◽  
Vol 230 (3) ◽  
pp. 797-805 ◽  
Author(s):  
M G Tordi ◽  
M C Silvestrini ◽  
A Colosimo ◽  
L Tuttobello ◽  
M Brunori
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Steady State ◽  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Binding Sites ◽  
Time Course ◽  
Product Inhibition ◽  
Inactive Form ◽  
Catalytically Active ◽  
The Stability

The kinetics of oxidation of azurin and cytochrome c-551 catalysed by Pseudomonas aeruginosa cytochrome oxidase were re-investigated, and the steady-state parameters were evaluated by parametric and non-parametric methods. At low concentrations of substrates (e.g. less than or equal to 50 microM) the values obtained for Km and catalytic-centre activity are respectively 15 +/- 3 microM and 77 +/- 6 min-1 for azurin and 2.15 +/- 0.23 microM and 66 +/- 2 min-1 for cytochrome c-551, in general accord with previous reports assigning to cytochrome c-551 the higher affinity for the enzyme and to azurin a slightly higher catalytic rate. However, when the cytochrome c-551 concentration was extended well beyond the value of Km, the initial velocity increased, and eventually almost doubled at a substrate concentration greater than or equal to 100 microM. This result suggests a ‘half-hearted’ behaviour, since at relatively low cytochrome c-551 concentrations only one of the two identical binding sites of the dimeric enzyme seems to be catalytically active, possibly because of unfavourable interactions influencing the stability of the Michaelis-Menten complex at the second site. When reduced azurin and cytochrome c-551 are simultaneously exposed to Ps. aeruginosa cytochrome oxidase, the observed steady-state oxidation kinetics are complex, as expected in view of the rapid electron transfer between cytochrome c-551 and azurin in the free state. In spite of this complexity, it seems likely that a mechanism involving a simple competition between the two substrates for the same active site on the enzyme is operative. Addition of a chemically modified and redox inactive form of azurin (Hg-azurin) had no effect on the initial rate of oxidation of either azurin and cytochrome c-551, but clearly altered the time course of the overall process by removing, at least partially, the product inhibition. The results lead to the following conclusions: (i) reduced azurin and cytochrome c-551 bind at the same site on the enzyme, and thus compete; (ii) Hg-azurin binds at a regulatory site, competing with the product rather than the substrate; (iii) the two binding sites on the dimeric enzyme, though intrinsically equivalent, display unfavourable interactions. Since water is the product of the reduction of oxygen, point (iii) has important implications for the reaction mechanism.

Kontakt- und Pseudokontakt-Verschiebungen in den 1H- und 13C-NMR-Spektren des paramagnetischen Komplexes Tricyclopentadienyl-(cyclohexylisonitril)-europium(III) / Contact- and Pseudocontact Shifts in the 1H- and 13C-NMR Spectra of the Paramagnetic, Organometallic Complex Tricyclopentadienyl-(cyclohexylisonitrile)-europium(III)

1973 ◽  
Vol 28 (3-4) ◽  
pp. 200-206 ◽  
Author(s):  
Reinhard Von Ammon ◽  
Basil Kanellakopulos ◽  
R. Dieter Fischer ◽  
Viktor Formacek
Keyword(s):  
13C Nmr ◽  
Shift Reagent ◽  
Positive Sign ◽  
Organometallic Complex ◽  
Pseudocontact Shifts ◽  
Europium Ion ◽  
Contact Shift ◽  
Polarization Mechanism ◽  
Negative Coupling ◽  
Paramagnetic Shifts

The organometallic NMR shift reagent Eu(C5H5)3 induces large paramagnetic shifts in the 1H-NMR spectrum of cyclohexylisonitrile. The 1:1-adduct (C5H5)3Eu•CNC6H11 has been isolated. The dipolar nature of the paramagnetic shifts of some of the C6H11-protons in this complex is used to fractionate the shifts of the C5H5-nuclei (1H and 13C) into the dipolar and contact contributions. The positive sign of the 13C contact shift is in accord with the usual spin alignment of the free europium ion relative to the external field together with a negative coupling constant. The opposite sign of the 1H contact shift is explained by a polarization mechanism. The anomalous temperature dependence of the 1H contact shift is connected with the strong second order paramagnetism of the europium ion.

scholarly journals Circular-dichroic properties and secondary structure of Pseudomonas aeruginosa soluble cytochrome c oxidase

1984 ◽  
Vol 218 (3) ◽  
pp. 907-912 ◽  
Author(s):  
M G Tordi ◽  
M C Silvestrini ◽  
A Colosimo ◽  
S Provencher ◽  
M Brunori
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Secondary Structure ◽  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Visible Region ◽  
Alpha Helix ◽  
Careful Analysis ◽  
Amino Acid Residues ◽  
Ferrocytochrome C ◽  
Significant Difference

The c.d. spectra of Pseudomonas aeruginosa cytochrome c oxidase in the oxidized state and the reduced state are reported in the visible- and u.v. absorption regions. In the visible region the comparison between the spectra of reduced cytochrome c oxidase and ferrocytochrome c-551 allows the identification of the c.d. bands mainly due to the d1 haem chromophore in cytochrome c oxidase. In the near-u.v. region the assignment of some of the observed peaks to the haem groups and to the aromatic amino acid residues is proposed. A careful analysis of the data in the far-u.v. region leads to the determination of the relative amounts of alpha-helix and beta-sheet in the enzyme, giving for the first time a picture of its secondary structure. A significant difference in this respect between the reduced and the oxidized species is observed and discussed in the light of similar conclusions reported by other workers.

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