Understanding of the binding interface between PsaC and the PsaA/PsaB heterodimer in photosystem I.
The PsaC subunit of Photosystem I (PS I) is tightly bound to the PsaA/PsaB heterodimer via an extensive network of ionic and hydrogen bonds. To improve our understanding of the design of the PsaC-PsaA/PsaB binding interface, variants of PsaC were generated, each lacking a key binding contact with the PsaA/PsaB heterodimer. The characteristics of the reconstituted, variant PS I complexes were monitored by time-resolved optical spectroscopy, low-temperature EPR spectroscopy, and electron transfer throughput measurements. In the absence of the ionic bond forming contacts R52(C) or R65(C), a markedly slower charge recombination occurs between P(700)(+) and [F(A)/F(B)](-). The addition of PsaD leads to the restoration of native recombination kinetics in a fraction of the PS I complexes reconstituted with R52A(C), but not with R65A(C). Contrary to expectation, the absence of Y80(C), which forms two symmetry-breaking H-bonds with PsaB, does not significantly affect the binding of PsaC as judged by the rate of charge recombination between P(700)(+) and [F(A)/F(B)](-). However, the removal of the entire C-terminus results in a dramatic decrease in the rate of charge recombination. Low-temperature EPR spectra of the variant PS I complexes indicate that the magnetic environments of F(A) and F(B) are altered when compared to that of native PS I. The slowing of the rate of charge recombination in the variant PS I complexes could be due to an increase in the distance between F(X) and F(A)/F(B) as the result of non-native binding or to an altered reduction potential of the iron-sulfur clusters, which would result in a different rate of thermalization up the electron acceptor chain. The most significant finding is that the variant PS I complexes support lower rates of light-induced flavodoxin reduction and that the rates deteriorate rapidly on exposure to dioxygen due to the degradation of F(A) and F(B). We suggest that the extensive set of ionic bonds and H-bonds between PsaC and the PsaA/PsaB heterodimer has evolved to ensure an exceedingly tight binding interface, thereby rendering the [4Fe-4S] clusters in PsaC inaccessible to dioxygen at the onset of oxygenic photosynthesis.
Department of Biochemistry and Molecular Biology, The Pennsylvania State University,University Park, Pennsylvania 16802, USA.
Bharat
B
Jagannathan
John H
JH
Golbeck
D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D006860:Hydrogen Bonding; D007700:Kinetics; D008958:Models, Molecular; D010100:Oxygen / Q000378:metabolism; D045331:Photosystem I Protein Complex / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D046940:Synechococcus / Q000378:metabolism; D013696:Temperature
D001426:Bacterial Proteins; D045331:Photosystem I Protein Complex; C089909:photosystem I, psaB subunit; D010100:Oxygen