A 19F nuclear magnetic resonance study of the interaction between cytochrome c and cytochrome c peroxidase.
The reaction between ferrocytochrome c and yeast cytochrome c peroxidase was studied using cytochrome c derivatives specifically trifluoroacetylated at single lysine amino groups. The only modifications that decreased the reaction rate were those of lysines immediately surrounding the heme crevice, lysines 13, 25, 79, and 87. Modification of lysines 22, 55, 88, and 99 had no effect on the reaction. The 19F chemical shifts of the cytochrome c derivatives trifluoroacetylated at lysines 13, 79, and 87 were not changed upon complex formation with cytochrome c peroxidase, indicating that no detectable conformational changes occurred. The cytochrome c trifluoroacetyl groups had the same T1 values in the paramagnetic fluorocytochrome c peroxidase complex as in the diamagnetic reduced form of the complex, indicating that they were more than 2.3 nm from the paramagnetic iron atom in cytochrome c peroxidase. This is consistent with a separation of at least 1.5-2.0 nm between the iron atom of cytochrome c and the iron atom of cytochrome c peroxidase.
D000818:Animals; D003574:Cytochrome c Group / Q000378:metabolism*; D003578:Cytochrome-c Peroxidase / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D006736:Horses; D009682:Magnetic Resonance Spectroscopy; D009206:Myocardium; D009994:Osmolar Concentration; D010544:Peroxidases / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation
D003574:Cytochrome c Group; D010544:Peroxidases; D003578:Cytochrome-c Peroxidase