Evidence that serine L223 is involved in the proton transfer pathway to QB in the photosynthetic reaction center of Rhodopseudomonas viridis.
In the reaction center of purple photosynthetic bacteria, the reducing equivalents produced by primary charge separation are exported via an ubiquinone molecule working as a two-electron shuttle. This loosely-bound quinone, called QB, accepts in successive flashes two electrons from the tightly bound primary quinone acceptor QA, along with two protons from the external medium. The surrounding protein plays an important role in stabilizing the semiquinone anion and in providing a pathway for protons from the cytoplasmic phase to QB. Herbicides of the triazine type compete with QB for the binding pocket and their binding is controlled by nearby amino acid residues. We have studied the kinetics of the first and second electron transfer from QA to QB in two herbicide-resistant mutants from Rhodopseudomonas viridis, T1 (ArgL217-->His,Ser L223-->Ala) and MAV5 (Arg L217-->His, Val L220-->Leu), in order to determine whether these residues are involved in proton transfer to the reduced QB. The main effect of the mutant T1 was a drastic (600-fold at pH 7) decrease in the rate of the second electron transfer to QB compared to the wild type. In contrast, the rate of the second electron transfer in the mutant MAV5 was decreased only slightly (10-fold) in the pH range from 7 to 11. We attribute the inhibition of the second electron transfer in the Ser L223-->Ala mutation to an essential role of Ser L223 in the donation of the first proton to the reduced QB.(ABSTRACT TRUNCATED AT 250 WORDS)
Leibl
W
W
Département de Biologie Cellulaire et Moléculaire, Centre d'Etudes de Saclay, Gif-sur-Yvette, France.
Sinning
I
I
Ewald
G
G
Michel
H
H
Breton
J
J
D000595:Amino Acid Sequence; D001665:Binding Sites; D004352:Drug Resistance, Microbial; D004579:Electron Transport; D006540:Herbicides / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D011487:Protein Conformation; D012241:Rhodopseudomonas / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D012694:Serine
D006540:Herbicides; D045322:Photosynthetic Reaction Center Complex Proteins; D012694:Serine