Study of QB- stabilization in herbicide-resistant mutants from the purple bacterium Rhodopseudomonas viridis.
The pH dependences of the rate constants of P+QB- (kBP) and P+QA- (kAP) charge recombination decays have been studied by flash-induced absorbance change technique, in chromatophores of three herbicide-resistant mutants from Rhodopseudomonas (Rps.) viridis, and compared to the wild type. P, QA, and QB are the primary electron donor and the primary and the secondary quinone acceptors, respectively. The triazine resistant mutants T1 (Arg L217----His and Ser L223----Ala), T3 (Phe L216----Ser and Val M263----Phe), and T4 (Tyr L222----Phe), all mutated in the QB binding pocket of the reaction center, have previously been characterized (Sinning, I., Michel, H., Mathis, P., & Rutherford, A. W. (1989) Biochemistry 28, 5544-5553). The pH dependence curves of kBP in T4 and the wild type are very close. This confirms that the sensitivity toward DCMU of T4 is mainly due to a structural rearrangement in the QB pocket rather than to a change in the charge distribution in this part of the protein. In T3, a 6-fold increase of kAP is observed (kAP = 4200 +/- 300 s-1 at pH 8) compared to that of the wild type (kAP = 720 +/- 50 s-1 at pH 8). We propose that the Val M263----Phe mutation induces a free energy decrease between P+QA- and P+I- (delta G zero IA) (I is the primary electron acceptor) of about 49 meV. The very different pH dependence of kAP in T3 suggests a substantial change in the QA pocket. The 2.5 times increase of kAP above pH 9.5 in the wild type is no longer detected in T3.(ABSTRACT TRUNCATED AT 250 WORDS)
Baciou
L
L
UPR 407, Bat 24 CNRS, Yvette, France.
Sinning
I
I
Sebban
P
P
D004352:Drug Resistance, Microbial / Q000235:genetics; D006540:Herbicides / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009154:Mutation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D011485:Protein Binding; D011809:Quinones / Q000737:chemistry*; D012241:Rhodopseudomonas / Q000187:drug effects / Q000235:genetics*; D013816:Thermodynamics
D006540:Herbicides; D045322:Photosynthetic Reaction Center Complex Proteins; D011809:Quinones