1QOB

FERREDOXIN MUTATION D62K

Summary for 1QOB

• Entry DOI: 10.2210/pdb1qob/pdb
• Descriptor: FERREDOXIN, SULFATE ION, FE2/S2 (INORGANIC) CLUSTER, ... (4 entities in total)
• Functional Keywords: electron transport, iron-sulfur, ferredoxin
• Biological source: Nostoc sp.
• Total number of polymer chains: 2
• Total formula weight: 21887.13

Authors

Holden, H.M.,Benning, M.M. (deposition date: 1997-08-14, release date: 1998-01-14, Last modification date: 2024-02-14)

Primary citation

Hurley, J.K.,Weber-Main, A.M.,Stankovich, M.T.,Benning, M.M.,Thoden, J.B.,Vanhooke, J.L.,Holden, H.M.,Chae, Y.K.,Xia, B.,Cheng, H.,Markley, J.L.,Martinez-Julvez, M.,Gomez-Moreno, C.,Schmeits, J.L.,Tollin, G.
Structure-function relationships in Anabaena ferredoxin: correlations between X-ray crystal structures, reduction potentials, and rate constants of electron transfer to ferredoxin:NADP+ reductase for site-specific ferredoxin mutants.
Biochemistry, 36:11100-11117, 1997

PubMed Abstract: A combination of structural, thermodynamic, and transient kinetic data on wild-type and mutant Anabaena vegetative cell ferredoxins has been used to investigate the nature of the protein-protein interactions leading to electron transfer from reduced ferredoxin to oxidized ferredoxin:NADP+ reductase (FNR). We have determined the reduction potentials of wild-type vegetative ferredoxin, heterocyst ferredoxin, and 12 site-specific mutants at seven surface residues of vegetative ferredoxin, as well as the one- and two-electron reduction potentials of FNR, both alone and in complexes with wild-type and three mutant ferredoxins. X-ray crystallographic structure determinations have been carried out for six of the ferredoxin mutants. None of the mutants showed significant structural changes in the immediate vicinity of the [2Fe-2S] cluster, despite large decreases in electron-transfer reactivity (for E94K and S47A) and sizable increases in reduction potential (80 mV for E94K and 47 mV for S47A). Furthermore, the relatively small changes in Calpha backbone atom positions which were observed in these mutants do not correlate with the kinetic and thermodynamic properties. In sharp contrast to the S47A mutant, S47T retains electron-transfer activity, and its reduction potential is 100 mV more negative than that of the S47A mutant, implicating the importance of the hydrogen bond which exists between the side chain hydroxyl group of S47 and the side chain carboxyl oxygen of E94. Other ferredoxin mutations that alter both reduction potential and electron-transfer reactivity are E94Q, F65A, and F65I, whereas D62K, D68K, Q70K, E94D, and F65Y have reduction potentials and electron-transfer reactivity that are similar to those of wild-type ferredoxin. In electrostatic complexes with recombinant FNR, three of the kinetically impaired ferredoxin mutants, as did wild-type ferredoxin, induced large (approximately 40 mV) positive shifts in the reduction potential of the flavoprotein, thereby making electron transfer thermodynamically feasible. On the basis of these observations, we conclude that nonconservative mutations of three critical residues (S47, F65, and E94) on the surface of ferredoxin have large parallel effects on both the reduction potential and the electron-transfer reactivity of the [2Fe-2S] cluster and that the reduction potential changes are not the principal factor governing electron-transfer reactivity. Rather, the kinetic properties are most likely controlled by the specific orientations of the proteins within the transient electron-transfer complex.

PubMed: 9287153
DOI: 10.1021/bi9709001

Experimental method

X-RAY DIFFRACTION (1.8 Å)