Plastocyanin cytochrome f interaction.
Spinach plastocyanin and turnip cytochrome f have been covalently linked by using a water-soluble carbodiimide to yield an adduct of the two proteins. The redox potential of cytochrome f in the adduct was shifted by -20 mV relative to that of free cytochrome f, while the redox potential of plastocyanin in the adduct was the same as that of free plastocyanin. Solvent perturbation studies showed the degree of heme exposure in the adduct to be less than in free cytochrome f, indicating that plastocyanin was linked in such a way as to bury the exposed heme edge. Small changes were also observed when the resonance Raman spectrum of the adduct was compared to that of free cytochrome f. The adduct was incapable of interacting with or donating electrons to photosystem I. Peptide mapping and sequencing studies revealed two sites of linkage between the two proteins. In one site of linkage, Asp-44 of plastocyanin is covalently linked to Lys-187 of cytochrome f. This represents the first identification of a group on cytochrome f that is involved in the interaction with plastocyanin. The other site of linkage involves Glu-59 and/or Glu-60 of plastocyanin to as yet unidentified amino groups on cytochrome f. Euglena cytochrome c-552 could also be covalently linked to turnip cytochrome f, although with a lower efficiency than spinach plastocyanin. In contrast, a variety of cyanobacterial cytochrome c-553's and a cyanobacterial plastocyanin could not be covalently linked to turnip cytochrome f.
Morand
L Z
LZ
Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville 72701.
Frame
M K
MK
Colvert
K K
KK
Johnson
D A
DA
Krogmann
D W
DW
Davis
D J
DJ
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D003580:Cytochromes / Q000032:analysis*; D045348:Cytochromes f; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000032:analysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010788:Photosynthesis; D010940:Plant Proteins / Q000032:analysis*; D010944:Plants / Q000201:enzymology*; D010970:Plastocyanin / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet; D013059:Spectrum Analysis, Raman
D003580:Cytochromes; D010940:Plant Proteins; D006418:Heme; D010970:Plastocyanin; D045348:Cytochromes f; D003488:Cyanogen Bromide