Herbicide binding and thermal stability of photosystem II isolated from Thermosynechococcus elongatus.
Binding of herbicides to photosystem II inhibits the electron transfer from Q(A) to Q(B) due to competition of herbicides with plastoquinone bound at the Q(B) site. We investigated herbicide binding to monomeric and dimeric photosystem II core complexes (PSIIcc) isolated from Thermosynechococcus elongatus by a combination of different methods (isothermal titration and differential scanning calorimetry, CD spectroscopy and measurements of the oxygen evolution) yielding binding constants, enthalpies and stoichiometries for various herbicides as well as information regarding stabilization/destabilization of the complex. Herbicide binding to detergent-solubilized PSIIcc can be described by a model of single independent binding sites present on this important membrane protein. Interestingly, binding stoichiometries herbicide:PSIIcc are lower than 1:1 and vary depending on the herbicide under study. Strong binding herbicides such as terbutryn stabilize PSIIcc in thermal unfolding experiments and endothermically binding herbicides like ioxynil probably cause large structural changes accompanied with the binding process as shown by differential scanning calorimetry experiments of the unfolding reaction of PSIIcc monomer in the presence of ioxynil. In addition we studied the occupancy of the Q(B) sites with plastoquinone (PQ9) by measuring flash induced fluorescence relaxation yielding a possible explanation for the deviations of herbicide binding from a 1:1 herbicide/binding site model.
Institute for Medical Physics and Biophysics, Charité-Universitätsmedizin, D-10098 Berlin, Schumann Str. 21/22, Germany.
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K
Zimmermann
M
M
Heck
J
J
Frank
J
J
Kern
I
I
Vass
A
A
Zouni
D001665:Binding Sites; D002151:Calorimetry / Q000379:methods; D000458:Cyanobacteria / Q000737:chemistry; D005453:Fluorescence; D006540:Herbicides / Q000737:chemistry*; D007462:Iodobenzenes / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D009570:Nitriles / Q000378:metabolism / Q000494:pharmacology; D045332:Photosystem II Protein Complex / Q000737:chemistry* / Q000187:drug effects; D010971:Plastoquinone / Q000032:analysis / Q000378:metabolism; D013816:Thermodynamics; D014227:Triazines / Q000378:metabolism / Q000494:pharmacology
D006540:Herbicides; D007462:Iodobenzenes; D009570:Nitriles; D045332:Photosystem II Protein Complex; D014227:Triazines; C008161:ioxynil; D010971:Plastoquinone; C010347:terbutryne