title: Equilibrium and kinetic parameters for the binding of inhibitors to the QB pocket in bacterial chromatophores: dependence on the state of QA.
abstract: The equilibrium and kinetic parameters for the binding of various inhibitors to the Q(B) pocket of the bacterial reaction center were investigated in chromatophores from Rhodobacter capsulatus and Rhodobacter sphaeroides. By monitoring the near-IR absorption changes specific to Q(A)(-) and Q(B)(-), we measured the fraction of inhibited centers in the dark and the kinetics and extent of inhibitor displacement after one flash due to the formation of the Q(A)Q(B)(-) state. The inhibitor release rate was much faster for triazines and o-phenanthroline (t(1/2) in the 50 ms to 1 s range) than for stigmatellin (t(1/2) approximately 20 s). For inhibitors with a rapid release rate, the fast phase of P(+) decay observed in the absence of secondary donor reflects the competition between P(+)Q(A)(-) recombination and inhibitor release: it is thus faster than the P(+)Q(A)(-) recombination, and its relative extent is smaller than the fraction of initially inhibited centers. At appropriate inhibitor concentrations, one can have almost total binding in the dark and almost total inhibitor displacement after one flash. Under such conditions, a pair of closely spaced flashes resets the two-electron gate in a single state (Q(A)Q(B)(-)), irrespective of the initial state. The apparent dissociation constant of terbutryn was significantly increased (by a factor of 4-7) in the presence of Q(A)(-), in agreement with the conclusion of Wraight and co-workers [Stein, R. R., et al. (1984) J. Cell. Biochem. 24, 243-259]. We suggest that this effect is essentially due to a tighter binding of ubiquinone in the Q(A)(-) state.
authors.affiliation: CEA-Cadarache, DSV-DEVM, Laboratoire de Bioénergétique Cellulaire, 13108 Saint-Paul-lez-Durance, France.
authors.forename: N
authors.initials: N
authors.lastname: Ginet
authors.forename: J
authors.initials: J
authors.lastname: Lavergne
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism; D016227:Benzoquinones / Q000737:chemistry / Q000378:metabolism; D020562:Betaproteobacteria / Q000187:drug effects / Q000378:metabolism; D001667:Binding, Competitive; D006540:Herbicides / Q000378:metabolism; D007700:Kinetics; D010782:Photolysis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D011090:Polyenes / Q000378:metabolism; D011809:Quinones / Q000737:chemistry / Q000378:metabolism*; D016354:Rhodobacter capsulatus / Q000187:drug effects / Q000378:metabolism; D012242:Rhodobacter sphaeroides / Q000187:drug effects / Q000378:metabolism; D012247:Rhodospirillum rubrum / Q000187:drug effects / Q000378:metabolism; D017186:Titrimetry; D014227:Triazines / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D016227:Benzoquinones; D006540:Herbicides; D045322:Photosynthetic Reaction Center Complex Proteins; D011090:Polyenes; D011809:Quinones; D014227:Triazines; C025232:semiquinone radicals; C041573:stigmatellin; C010347:terbutryne