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PMID: 1756713 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The surface-exposed tyrosine residue Tyr83 of pea plastocyanin is involved in both binding and electron transfer reactions with cytochrome f.

The EMBO journal ·Vol. 10 ·No. 13 ·1991-12-00 ·Pages 4011-6

He S, Modi S, Bendall DS, Gray JC

Abstract

Site-directed mutants of the pea plastocyanin gene in which the codon for the surface-exposed Tyr83 has been changed to codons for Phe83 and Leu83 have been expressed in transgenic tobacco plants. The mutant proteins have been purified to homogeneity and their conformations shown not to differ significantly from the wild-type plastocyanin by 1H-NMR and CD. Overall rate constants for electron transfer (k2) from cytochrome f to plastocyanin have been measured by stopped-flow spectrophotometry and rate constants for binding (ka) and association constants (KA) have been measured from the enhanced Soret absorption of cytochrome f on binding plastocyanin. These measurements allow the calculation of the intrinsic rate of electron transfer in the binary complex. An 8-fold decrease in the overall rate of electron transfer to the Phe83 mutant is due entirely to a decreased association constant for cytochrome f, whereas the 40-fold decrease in the overall rate of electron transfer to the Leu83 mutant is due to weaker binding and a lower intrinsic rate of electron transfer. This indicates that Tyr83 is involved in binding to cytochrome f and forms part of the main route of electron transfer.

MeSH Terms
Base Sequence Binding Sites Circular Dichroism Cytochromes/metabolism Cytochromes f Deoxyribonucleotides Electrons Fabaceae/genetics,metabolism Kinetics Magnetic Resonance Spectroscopy Molecular Sequence Data Mutagenesis, Site-Directed Plants, Genetically Modified/genetics Plants, Medicinal Plasmids Plastocyanin/genetics,metabolism Restriction Mapping Spectrophotometry, Ultraviolet Tyrosine/genetics
Chemicals
Cytochromes Deoxyribonucleotides Tyrosine Plastocyanin Cytochromes f
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
He S
Cambridge Centre for Molecular Recognition, University of Cambridge, UK.
Modi S
Bendall D S
Gray J C
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1991-12-00
Pages
4011-6
Language
English
Region
England
NLM ID
8208664
PMCID
PMC453148
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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