PDBsum entry 1j7a

Electron transport

PDB id: 1j7a

Contents

Protein chain

98 a.a. *

Ligands

FES

Waters ×100

* Residue conservation analysis

PDB id:

1j7a

Name:

Electron transport

Title:

Structure of the anabaena ferredoxin d68k mutant

Structure:

Ferredoxin i. Chain: a. Engineered: yes. Mutation: yes

Source:

Nostoc sp. Pcc. Organism_taxid: 103690. Strain: 7120. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.80Å R-factor: 0.174

Authors:

J.K.Hurley,A.M.Weber-Main,M.T.Stankovich,M.M.Benning,J.B.Thoden, J.L.Vanhooke,H.M.Holden,Y.K.Chae,B.Xia,H.Cheng,J.L.Markley, M.Martinez-Julvez,C.Gomez-Moreno,J.L.Schmeits,G.Tollen

Key ref:

J.K.Hurley et al. (1997). 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. PubMed id: 9287153 DOI: 10.1021/bi9709001

Date:

16-May-01 Release date: 23-May-01

Supersedes:

1qoc

Protein chain

P0A3C7  (FER1_NOSS1) -  Ferredoxin-1 from Nostoc sp. (strain PCC 7120 / SAG 25.82 / UTEX 2576)

Seq: 99 a.a.

Struc: 98 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1021/bi9709001

Biochemistry 36:11100-11117 (1997)

PubMed id: 9287153

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.

J.K.Hurley, A.M.Weber-Main, M.T.Stankovich, M.M.Benning, J.B.Thoden, J.L.Vanhooke, H.M.Holden, Y.K.Chae, B.Xia, H.Cheng, J.L.Markley, M.Martinez-Júlvez, C.Gómez-Moreno, J.L.Schmeits, G.Tollin.

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.