Transient kinetics of electron transfer from a variety of c-type cytochromes to plastocyanin.
Plastocyanin (PC) and its physiological reaction partner cytochrome (cyt) f form a complex which is electrostatically stabilized by interactions between complementary localized charges. We have measured the kinetics of intracomplex electron transfer between several reduced cytochromes and PC using laser flash photolysis. With spinach cyt f and spinach PC, we obtain first-order rate constants, kforward = 2780 s-1 and kreverse = 1050 s-1, for the reversible reaction and a complex dissociation constant of about 23 microM at an ionic strength (I) of 5 mM. The observed rate constant increases by a factor of 2 between I = 5 and 40 mM and then decreases monotonically at higher ionic strengths. This indicates that the complex is not completely dissociated until I = 150 mM and that the proteins within the electrostatically most stable complex are not optimally oriented for electron transfer. Similar results were obtained with turnip cyt f and spinach PC, although in this case intracomplex electron transfer is about 4 times as fast. Horse cyt c also forms an electrostatically stabilized complex with PC, and yields a limiting rate constant for intracomplex electron transfer (1750 s-1) and a dissociation constant (10 microM) comparable to those for spinach cyt f. The ionic strength dependence shows that the complex is more readily dissociated (complete at I = 25 mM) than is that of cyt f and that rearrangement is not required for optimal electron transfer. Addition of polylysine results in 10-fold inhibition of the rate of electron transfer. Pseudomonas cyt c-551 is an acidic cytochrome which does not form a complex with PC.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Biochemistry, University of Arizona, Tucson 85721.
T E
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Meyer
Z G
ZG
Zhao
M A
MA
Cusanovich
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G
Tollin
D001426:Bacterial Proteins; D003574:Cytochrome c Group / Q000378:metabolism*; D004563:Electrochemistry; D004579:Electron Transport; D007700:Kinetics; D007834:Lasers; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D010782:Photolysis; D010944:Plants / Q000737:chemistry; D010970:Plastocyanin / Q000378:metabolism*; D011107:Polylysine / Q000494:pharmacology
D001426:Bacterial Proteins; D003574:Cytochrome c Group; D011107:Polylysine; D010970:Plastocyanin; C017167:cytochrome C(551); C018887:cytochrome c-555