Purification and properties of cytochrome c (550; Hemispora stellata)

1973 ◽  
Vol 88 (1) ◽  
pp. 11-24 ◽  
Author(s):  
J. Francisco T. Sala ◽  
Justino Burgos
Keyword(s):  
Cytochrome C ◽  
Purification And Properties

scholarly journals Purification and properties of a cross-linked complex between cytochrome c and cytochrome c peroxidase

1982 ◽  
Vol 201 (1) ◽  
pp. 9-18 ◽  
Author(s):  
G W Pettigrew ◽  
S Seilman
Keyword(s):  
Cytochrome C ◽  
Cytochrome C Peroxidase ◽  
Sodium Phosphate Buffer ◽  
Oxidation Reduction ◽  
Purification And Properties ◽  
Lysine Residues ◽  
The Cross ◽  
Nadh Cytochrome ◽  
Stable Association ◽  
Covalently Linked

Cytochrome c (horse heart) was covalently linked to yeast cytochrome c peroxidase by using the cleavable bifunctional reagent dithiobis-succinimidyl propionate in 5 mM-sodium phosphate buffer, pH 7.0. A cross-linked complex of molecular weight 48 000 was purified in approx. 10% yield from the reaction mixture, which contained 1 mol of cytochrome c and 1 mol of cytochrome c peroxidase/mol. Of the total 40 lysine residues, four to six were blocked by the cross-linking agent. Dithiobis-succinimidylpropionate can also cross-link cytochrome c to ovalbumin, but cytochrome c peroxidase is the preferred partner for cytochrome c in a mixture of the three proteins. The cytochrome c cross-linked to the peroxidase can be rapidly reduced by free cytochrome c-557 from Crithidia oncopelti, and the equilibrium obtained can be used to calculate a mid-point oxidation-reduction potential for the cross-linked cytochrome of 243 mV. Mitochondrial NADH-cytochrome c reductase will reduce the bound cytochrome only very slowly, but the rate of reduction by ascorbate at high ionic strength approaches that for free cytochrome c. Bound cytochrome c reduced by ascorbate can be re-oxidized within 10s by the associated peroxidase in the presence of equimolar H2O2. In the standard peroxidase assay the cross-linked complex shows 40% of the activity of the free peroxidase. Thus the intrinsic ability of each partner in the complex to take part in electron transfer is retained, but the stable association of the two proteins affects access of reductants.

Purification and Properties of Cytochrome C-553 from the Green Sulfur Bacterium Chlorobium Tepidum

1998 ◽  
pp. 1593-1596 ◽  
Author(s):  
Fabiyola Selvaraj ◽  
Dominica Devine ◽  
Wenli Zhou ◽  
Daniel C. Brune ◽  
Michael T. Lince ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Chlorobium Tepidum ◽  
Green Sulfur Bacterium ◽  
Purification And Properties ◽  
Green Sulfur

Human cytochrome c oxidase isoenzymes from heart and skeletal muscle; purification and properties

1987 ◽  
Vol 890 (2) ◽  
pp. 144-150 ◽  
Author(s):  
Karin M.C. Sinjorgo ◽  
Theo B.M. Hakvoort ◽  
Ilker Durak ◽  
J.Wim Draijer ◽  
Jan K.P. Post ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Human Cytochrome ◽  
Purification And Properties ◽  
Human Cytochrome C

scholarly journals Solubilization, partial purification and properties of N-methylglutamate dehydrogenase from Pseudomonas aminovorans

1977 ◽  
Vol 161 (2) ◽  
pp. 357-370 ◽  
Author(s):  
C W Bamforth ◽  
P J Large
Keyword(s):  
Cytochrome C ◽  
Electron Acceptor ◽  
Specific Activity ◽  
Partial Purification ◽  
Ph Optimum ◽  
Transport Chain ◽  
Purification And Properties ◽  
Solubilized Enzyme ◽  
Triton X ◽  
Sepharose 4B

1. Extracts of amine-grown Pseudomonas aminovorans contained a particle-bound N-methylglutamate dehydrogenase (EC 1.5.99.5). The enzyme was not present in succinate-grown cells, and activity appeared before growth began in succinate-grown cells which had been transferred to methylamine growth medium. 2. Membrane-containing preparations from methylamine-grown cells catalysed an N-methylglutamate-dependent uptake of O2 or reduction of cytochrome c, which was sensitive to inhibitors of the electron-transport chain. 3. N-Methylglutamate dehydrogenase activity with phenazine methosulphate or 2,6-dichlorophenol-indophenol as electron acceptor could be solubilized with 1% (w/v) Triton X-100. The solubilized enzyme was much less active with cytochrome c as electron acceptor and did not sediment in 1 h at 150000g. Solubilization was accompanied by a change in the pH optimum for activity. 4. The solubilized enzyme was partially purified by Sepharose 4B and hydroxyapatite chromatograpy to yield a preparation 22-fold increased in specific activity over the crude extract. 5. The partially-purified enzyme was active with sarcosine, N-methylalanine and N-methylaspartate as well as with N-methylglutamate. Evidence suggesting activity with N-methyl D-amino acids as well as with the L-forms was obtained. 6. The enzyme was inhibited by p-chloromercuribenzoate, iodoacetamide and by both ionic and non-ionic detergents. 2-Oxoglutarate and formaldehyde were also inhibitors. 7. Kinetic analysis confirmed previous workers' observations of a group transfer (Ping Pong) mechanism. 8. Spectral observations suggested that the partially purified preparation contained flavoprotein and a b-type cytochrome. 9. The role of the enzyme in the oxidation of methylamine is discussed.

scholarly journals Purification and properties of cytochrome c

1945 ◽  
Vol 39 (4) ◽  
pp. 289-292 ◽  
Author(s):  
D. Keilin ◽  
E. F. Hartree
Keyword(s):  
Cytochrome C ◽  
Purification And Properties
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