Diphenyl Ether Herbicides, a Tool to Elucidate the Mechanism of Photophosphorylation

At least two different classes of ADP binding sites on chloroplast coupling factor are de­ scribed in the literature. High-affinity sites are assumed to entail regulatory functions while low-affinity sites are involved in catalysis. Diphenyl ether herbicides, acting as energy transfer inhibitors, interfere with nucleotide ex­ change on both categories of ADP binding sites. Their inhibitory potency varies depending on their substitution. We found that each diphenyl ether assayed revealed identical / 50 values for inhibition of nucleotide interaction with both classes of binding sites. We show here that diphenyl ether inhibition of energytransfer is primary based upon the interference with ADP binding to high-affinity binding sites. Thereby the control of proton permeability through the coupling factor complex is affected. Moreover, we found that the three ß-subunits are not absolutely fixed in one conformational state: After covalently block­ ing the high-affinity site by an azido-label, a new ß-subunit.