Role of the hydrogen bond from Leu722 to the A1A phylloquinone in photosystem I.
Photosystem I (PS I) contains two molecules of phylloquinone that function as electron transfer cofactors at highly reducing midpoint potentials. It is therefore surprising that each phylloquinone is hydrogen bonded at the C(4) position to the backbone -NH of a Leu residue since this serves to drive the midpoint potential more oxidizing. To better understand the role of the H-bond, a PS I variant was generated in which L722(PsaA) was replaced with a bulky Trp residue. This change was designed to alter the conformation of the A-jk(1) loop and therefore change the strength of the H-bond to the PsaA-branch phylloquinone. Transient EPR studies at 80 K show that the A(1A) site in the PS I variant is fully occupied with phylloquinone, but the absence of methyl hyperfine couplings in the quinone contribution to the P(700)(*+)A(1)(*-) radical pair spectrum indicates that the H-bond has been weakened. In wild-type PS I, reduction of F(A) and F(B) with sodium dithionite causes a approximately 30% increase in the amplitude of the P(700)(*+)A(1)(*-) transient EPR signal due to the added contribution of the PsaB-branch cofactors to low temperature reversible electron transfer between P(700) and A(1A). In contrast, the same treatment to the L722W(PsaA) variant leads to a approximately 75% reduction in the amplitude of the P(700)(*+)A(1)(*-) transient EPR signal. This behavior suggests that A(1A) has undergone double reduction to phyllohydroquinone, thereby preventing electron transfer past A(0A). The remaining 25% of the P(700)(*+)A(1)(*-) radical pair spectrum shows an altered spin polarization pattern and pronounced methyl hyperfine couplings characteristic of asymmetric H-bonding to the phylloquinone. Numerical simulations of the polarization pattern indicate that it arises primarily from electron transfer between P(700) and A(1B). The altered reduction behavior in the L722W(PsaA) variant suggests that the primary purpose of the H-bond is to tie up the C(4) carbonyl group of phylloquinone in a H-bond so as to prevent protonation and hence lower the probability of double reduction during periods of high light intensity.
Srinivasan
Nithya
N
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
Karyagina
Irina
I
Bittl
Robert
R
van der Est
Art
A
Golbeck
John H
JH
D019943:Amino Acid Substitution / Q000235:genetics; D018360:Crystallography, X-Ray; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals / Q000737:chemistry; D006860:Hydrogen Bonding; D007930:Leucine / Q000737:chemistry* / Q000235:genetics; D010084:Oxidation-Reduction; D045331:Photosystem I Protein Complex / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D017354:Point Mutation; D046939:Synechocystis / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D013696:Temperature; D014364:Tryptophan / Q000235:genetics; D010837:Vitamin K 1 / Q000737:chemistry* / Q000378:metabolism
D005609:Free Radicals; D045331:Photosystem I Protein Complex; D010837:Vitamin K 1; D014364:Tryptophan; D007930:Leucine