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Slow formation of [3Fe–4S] 1+ clusters in mutant forms of Desulfovibrio africanus ferredoxin III
Author(s) -
Hannan Jonathan P.,
Busch Johanneke L.H.,
James Richard,
Thomson Andrew J.,
Moore Geoffrey R.,
Davy Sharon L.
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)01210-2
Subject(s) - ferredoxin , mutant , desulfovibrio , chemistry , crystallography , biology , bacteria , biochemistry , genetics , gene , enzyme
Desulfovibrio africanus ferredoxin III ( Da FdIII) readily interconverts between a 7Fe and an 8Fe form with Asp‐14 believed to provide a cluster ligand in the latter form. To investigate the factors important for cluster interconversion in Fe/S cluster‐containing proteins we have studied two variants of Da FdIII produced by site‐directed mutagenesis, Asp14Glu and Asp14His, with cluster incorporation performed in vitro. Characterisation of these proteins by UV/visible, EPR and 1 H NMR spectroscopies revealed that the formation of the stable 7Fe form of these proteins takes some time to occur. Evidence is presented which indicates the [4Fe–4S] 2+ cluster is incorporated prior to the [3Fe–4S] 1+ cluster.