[Pyridoxalphosphate-modified derivatives of cytochrome c. Mono- and disubstituted derivatives: characteristics and effect on electron transport in cytochrome c-depleted mitochondria].
Cytochrome c is modified by covalent binding of pyridoxal phosphate (PLP) to lysine residues. One di-substituted [(PLP)2--C] and two mono-substituted derivatives [(PLP)--c and (PLP)''--c] were obtained and precisely purified. The peak at 695 nm and CD-spectra in 190--600 nm region show that all derivatives have native conformation. The differential UV-spectra of the derivatives against native protein show that in (PLP)2--c there is a contact dipole-dipole interaction between PLP chromophores. It is calculated that the N-atoms of the two PLP-substituted lysines must be at a distance less than or equal to 12 A. Analysing our and literature data, one may suppose that Lys-13 and Lys-87 are the most probable candidates for modification with PLP. (PLP)---c and (PLP)''--c behave differently during ion-exchange chromatography and when added to cytochrom c-depleted mitochondria. (PLP)''--c restores electron transfer at higher concentrations than (PLP)'--c. Both they restore fully succinate and ascorbate oxidation but at considerably higher concentrations than the native protein, i. e. modification of any one of the reactive towards PLP lysines descreases but does not exclude the interaction with its reductase and oxidase. The effective equilibrium constants of binding of modified derivatives to cytochrome c-depleted mitochondria are lower than the constant for native protein. Together with decrease in binding activity, Hill coefficients increase. From our results it may be supposed that probably the binding sites of cytochrome c for its reductase and oxidase partially overlap.
D000818:Animals; D001665:Binding Sites; D003574:Cytochrome c Group / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D004579:Electron Transport / Q000187:drug effects*; D066298:In Vitro Techniques; D008239:Lysine / Q000378:metabolism; D008930:Mitochondria, Liver / Q000378:metabolism*; D008968:Molecular Conformation; D010084:Oxidation-Reduction / Q000187:drug effects; D011732:Pyridoxal Phosphate; D051381:Rats; D013057:Spectrum Analysis; D013268:Stimulation, Chemical; D013816:Thermodynamics
D003574:Cytochrome c Group; D011732:Pyridoxal Phosphate; D008239:Lysine
Modifitsirovannye piridoksal'fosfatom proizvodnye tsitokhroma c. Mono- i dizameshchennye proizvodnye: kharakteristika i vliianie na perenos élektronov v obednennykh tsitokhromom c mitokhondriiakh.