Interaction of an engineered [3Fe-4S] cluster with a menaquinol binding site of Escherichia coli DMSO reductase

Biochemistry
Richard A Rothery, J H Weiner

Abstract

We have characterized by EPR the interaction of the Em,7 = -50 mV [4Fe-4S] cluster of Escherichia coli DMSO reductase (DmsABC) with a menaquinol (MQH2) binding site. Potentiometric titrations indicate that in DmsAB(C102S)C, the Em,7 = -50 mV [4Fe-4S] cluster is replaced by an Em,7 = +260 mV [3Fe-4S] cluster. The Q-pool coupling assay in combination with the MQH2 analog HOQNO (2-n-heptyl-4-hydroxyquinoline-N-oxide) was used to examine the effect of the DmsB(Cl02S) mutation on physiological electron transfer through DmsABC. Forward electron transfer through the mutant (MQH2 to DmsA) is blocked in the Q-pool coupling assay, but reverse electron transfer (DmsA to MQ) is not. HOQNO elicits a significant change in the EPR line shape of the oxidized DmsAB(Cl02S)C [3Fe-4S] cluster but has no effect on the line shape of the reduced [4Fe-4S] clusters. We have identified a residue in DmsC involved in MQH2 oxidation. DmsC(H65), and in a double mutant, DmsAB(C102S)C(H65R), the DmsC mutation blocks the HOQNO effect on the [3Fe-4S] EPR line shape, suggesting, that the DmsC(H65R) mutation either blocks HOQNO binding or blocks a conformational link between a HOQNO binding site and the DmsB(C102S) [3Fe-4S] cluster. These results suggest that the...Continue Reading

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Citations

Apr 4, 2002·Molecular Microbiology·Nicola R StanleyBen C Berks
May 26, 2010·The Journal of Biological Chemistry·Sofia S VenceslauInes A C Pereira
Sep 18, 2002·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Damaraju SambasivaraoJoel H Weiner
Jan 29, 2014·Chemical Reviews·Russ HillePartha Basu
Jan 16, 2013·Dalton Transactions : an International Journal of Inorganic Chemistry·Russ Hille
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Oct 3, 2003·Archives of Biochemistry and Biophysics·Shannon P Lubitz, Joel H Weiner
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Feb 6, 2004·Biochimica Et Biophysica Acta·Paulina Geijer, Joel H Weiner
Oct 30, 2007·Biochimica Et Biophysica Acta·Richard A RotheryJoel H Weiner
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Apr 1, 2007·EcoSal Plus·Victor W T Cheng, Joel H Weiner
Nov 7, 1996·Chemical Reviews·Russ Hille

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