Some reactions of carbon monoxide and oxygen with carbodi-imide-modified cytochrome c.
The reactivity of carbodi-imide-modified tuna and horse heart cytochromes c with the ferrous ion ligands CO and O2 has been studied. Both modified cytochromes bind one molecule of CO. Stopped-flow and flash-photolysis experiments indicate the presence of three kinetic processes in the reaction of the cytochromes with CO. The second-order rate constants associated with all three kinetic process are pH-independent being 2.8 x 10(5) M-1.s-1, 3.8 x 10(4) M-1.s-1 and 4 x 10(3) M-1.s-1 under all conditions studied. The concentration-dependence of the contributions made by each of the processes to the overall absorbance change indicates that the fast and slow kinetic phases are associated with two forms of the cytochromes which are in equilibrium, whereas the intermediate phase arises from a separate cytochrome species. The quantum yield for the photodissociation of CO from the ferrous cytochromes is unusually low. Both modified cytochromes are capable of binding and reducing O2. In the presence of excess reductant, the modified cytochromes can catalytically reduce large molar excesses of O2. In the absence of excess reducing agent, the oxy complex initially formed undergoes a pH-dependent intramolecular electron-transfer process with half-life approx. 10 min. EDC [1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide]-promoted internal cross-linking is proposed to account for differences between the EDC-modified proteins and carboxymethylated cytochrome c.
Mathews
A J
AJ
Department of Biochemistry, University of Auckland, New Zealand.
Brittain
T
T
D000818:Animals; D002248:Carbon Monoxide / Q000378:metabolism* / Q000494:pharmacology; D003574:Cytochrome c Group / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000494:pharmacology*; D006736:Horses; D007700:Kinetics; D009206:Myocardium / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D010782:Photolysis; D011485:Protein Binding; D013053:Spectrophotometry; D014413:Tuna
D003574:Cytochrome c Group; D002248:Carbon Monoxide; D005022:Ethyldimethylaminopropyl Carbodiimide; D010100:Oxygen