2b7r
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Structure of E378D mutant flavocytochrome c3
OverviewOverview
The mechanism for fumarate reduction by the soluble fumarate reductase, from Shewanella frigidimarina involves hydride transfer from FAD and, proton transfer from the active-site acid, Arg-402. It has been proposed, that Arg-402 forms part of a proton transfer pathway that also involves, Glu-378 and Arg-381 but, unusually, does not involve any bound water, molecules. To gain further insight into the importance of this proton, pathway we have perturbed it by substituting Arg-381 by lysine and, methionine and Glu-378 by aspartate. Although all the mutant enzymes, retain measurable activities, there are orders-of-magnitude decreases in, their k(cat) values compared with the wild-type enzyme. Solvent kinetic, isotope effects show that proton transfer is rate-limiting in the, wild-type and mutant enzymes. Proton inventories indicate that the proton, pathway involves multiple exchangeable groups. Fast scan protein-film, voltammetric studies on wild-type and R381K enzymes show that the proton, transfer pathway delivers one proton per catalytic cycle and is not, required for transporting the other proton, which transfers as a hydride, from the reduced, protonated FAD. The crystal structures of E378D and, R381M mutant enzymes have been determined to 1.7 and 2.1 A resolution, respectively. They allow an examination of the structural changes that, disturb proton transport. Taken together, the results indicate that, Arg-381, Glu-378, and Arg-402 form a proton pathway that is completely, conserved throughout the fumarate reductase/succinate dehydrogenase family, of enzymes.
About this StructureAbout this Structure
2B7R is a Single protein structure of sequence from Shewanella frigidimarina with NA, HEM, FAD and FUM as ligands. Active as Succinate dehydrogenase, with EC number 1.3.99.1 Full crystallographic information is available from OCA.
ReferenceReference
A proton delivery pathway in the soluble fumarate reductase from Shewanella frigidimarina., Pankhurst KL, Mowat CG, Rothery EL, Hudson JM, Jones AK, Miles CS, Walkinshaw MD, Armstrong FA, Reid GA, Chapman SK, J Biol Chem. 2006 Jul 21;281(29):20589-97. Epub 2006 May 12. PMID:16699170
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