Identification of the proton pathway in bacterial reaction centers: replacement of Asp-M17 and Asp-L210 with asn reduces the proton transfer rate in the presence of Cd2+.
The reaction center (RC) from Rhodobacter sphaeroides converts light into chemical energy through the reduction and protonation of a bound quinone molecule Q(B) (the secondary quinone electron acceptor). We investigated the proton transfer pathway by measuring the proton-coupled electron transfer, k(AB)((2)) [Q(A)Q(B) + H(+) --> Q(A)(Q(B)H)(-)] in native and mutant RCs in the absence and presence of Cd(2+). Previous work has shown that the binding of Cd(2+) decreases k(AB)((2)) in native RCs approximately 100-fold. The preceding paper shows that bound Cd(2+) binds to Asp-H124, His-H126, and His-H128. This region represents the entry point for protons. In this work we investigated the proton transfer pathway connecting the entry point with Q(B) by searching for mutations that greatly affect k(AB)((2)) ( greater, similar10-fold) in the presence of Cd(2+), where k(AB)((2)) is limited by the proton transfer rate (k(H)). Upon mutation of Asp-L210 or Asp-M17 to Asn, k(H) decreased from approximately 60 s(-1) to approximately 7 s(-1), which shows the important role that Asp-L210 and Asp-M17 play in the proton transfer chain. By comparing the rate of proton transfer in the mutants (k(H) approximately 7 s(-1)) with that in native RCs in the absence of Cd(2+) (k(H) >/= 10(4) s(-1)), we conclude that alternate proton transfer pathways, which have been postulated, are at least 10(3)-fold less effective.
Paddock
M L
ML
Department of Physics 0319, 9500 Gilman Drive, University of California at San Diego, La Jolla, CA 92093, USA.
Feher
G
G
Okamura
M Y
MY
D001224:Aspartic Acid / Q000737:chemistry / Q000378:metabolism*; D016227:Benzoquinones / Q000378:metabolism; D002104:Cadmium / Q000494:pharmacology*; D004579:Electron Transport; D007700:Kinetics; D007834:Lasers; D008958:Models, Molecular; D009154:Mutation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics*; D011485:Protein Binding; D011522:Protons; D011809:Quinones / Q000378:metabolism; D012242:Rhodobacter sphaeroides
D016227:Benzoquinones; D045322:Photosynthetic Reaction Center Complex Proteins; D011522:Protons; D011809:Quinones; D002104:Cadmium; D001224:Aspartic Acid; C025232:semiquinone radicals