Structure of C73G putidaredoxin from Pseudomonas putida

Journal Article · Thu Jan 01 04:00:00 EST 2004 · Acta Crystallographica Section D: Biological Crystallography

DOI:https://doi.org/10.1107/S0907444904003348· OSTI ID:930072

Smith, N; Mayhew, M; Holden, M; Kelly, H; Robinson, H; Heroux, A; Vilker, V; Gallagher, D

The structure of the C73G mutant of putidaredoxin (Pdx), the Fe{sub 2}S{sub 2} ferredoxin that supplies electrons to cytochrome CYP101 (P450cam) for camphor oxidation, is reported at 1.9 {angstrom} resolution in a C2 crystal form. The structure was solved by single-wavelength iron anomalous diffraction, which yielded electron density above the 2{sigma} level for over 97% of the non-H atoms in the protein. The final structure with R = 0.19 and R{sub free} = 0.21 has been deposited in the Protein Data Bank with accession code 1r7s. The C2 crystal contains three Pdx molecules in the asymmetric unit, giving three independent models of the protein that are very similar (r.m.s.d. < 0.3 {angstrom} for the 106 C{sup {alpha}} atoms). The unusually high solvent fraction of 80% results in comparatively few crystal-packing artifacts. The structure is briefly compared with the recently reported crystal structures of the C73S and C73S/C85S mutants. In general, the eight independent molecules in the three crystal structures (three in C73G, three in C73S and two in C73S/C85S) are much more similar to each other than to the previously reported NMR structure of wild-type Pdx in solution. The present findings show a unanimous structure in some regions crucial for electron-transfer interactions, including the cluster-binding loop 39-48 and the cytochrome-interaction region of Asp38 and Trp106. In addition, the Cys45 amide group donates a hydrogen bond to cluster sulfur S1, with Ala46 adopting an L conformation, in all three molecules in the crystal.

• Research Organization: Brookhaven National Laboratory (BNL) National Synchrotron Light Source
• Sponsoring Organization: Doe - Office Of Science
• DOE Contract Number: AC02-98CH10886
• OSTI ID: 930072
• Report Number(s): BNL--80701-2008-JA
• Journal Information: Acta Crystallographica Section D: Biological Crystallography, Journal Name: Acta Crystallographica Section D: Biological Crystallography Vol. 60
• Country of Publication: United States
• Language: English

Related Subjects

08 HYDROGEN
32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATION
36 MATERIALS SCIENCE
AMIDES
ATOMS
CAMPHOR
COMMERCIAL BUILDINGS
CRYSTAL STRUCTURE
CRYSTALS
CYTOCHROMES
DATA
DIFFRACTION
ELECTRON DENSITY
ELECTRON TRANSFER
ELECTRONS
FERREDOXIN
HYDROGEN
INTERACTIONS
IRON
LEVELS
MOLECULES
MUTANTS
NUCLEAR MAGNETIC RESONANCE
OXIDATION
PROTEINS
PSEUDOMONAS
RESOLUTION
SOLVENTS
SULFUR
national synchrotron light source