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The Catalytic Subunit of Escherichia coli Nitrate Reductase A ...

The Catalytic Subunit of Escherichia coli Nitrate Reductase A ...

The Catalytic Subunit of Escherichia coli Nitrate Reductase A

5324 Biochemistry 2004, 43, 5324-5333The Catalytic Subunit of Escherichia coli Nitrate Reductase A Contains a Novel[4Fe-4S] Cluster with a High-Spin Ground State †,‡Richard A. Rothery, § Michela G. Bertero, | Richard Cammack, ⊥ Monica Palak, § Francis Blasco, #Natalie C. J. Strynadka, | and Joel H. Weiner* ,§CIHR Membrane Protein Research Group, Department of Biochemistry, UniVersity of Alberta, 474 Medical Sciences Building,Edmonton, Alberta T6G 2H7, Canada, Department of Biochemistry, UniVersity of British Columbia, 2146 Health Sciences Mall,VancouVer, British Columbia V6T 1Z3, Canada, Department of Life Sciences, King’s College London,Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NN, U.K., and Laboratoire de Chimie Bactérienne,CNRS, 31 chemin Joseph Aiguier, 13402 Marseille Cedex 9, FranceReceiVed January 8, 2004; ReVised Manuscript ReceiVed March 3, 2004ABSTRACT: We have used EPR spectroscopy, redox potentiometry, and protein crystallography tocharacterize the [4Fe-4S] cluster (FS0) of the Escherichia coli nitrate reductase A (NarGHI) catalyticsubunit (NarG). FS0 is clearly visible in the crystal structure of NarGHI [Bertero, M. G., et al. (2003)Nat. Struct. Biol. 10, 681-687] but has novel coordination comprising one His residue and three Cysresidues. At low temperatures (

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