A pure S=3/2 [Fe4S4]+ cluster in the A33Y variant of Pyrococcus furiosus ferredoxin

FEBS Letters

Johnson, Michael K¹  Duderstadt, Randall E¹  Brereton, Phillip S¹  Adams, Michael W.W¹ 

[1] Departments of Chemistry, and Biochemistry and Molecular Biology, and the Center for Metalloenzyme Studies, University of Georgia, Athens, GA 30602, USA

关键词: Ferredoxin;    Iron-sulfur cluster;    Pyrococcus furiosus;    EPR;    Magnetic circular dichroism;    Resonance Raman;   

DOI  :  10.1016/S0014-5793(99)00766-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The properties of the [4Fe-4S]^2+/+ cluster in wild-type and the A33Y variant of Pyrococcus furiosus ferredoxin have been investigated by the combination of EPR, variable-temperature magnetic circular dichroism (VTMCD) and resonance Raman (RR) spectroscopies. The A33Y variant involves the replacement of an alanine whose α-C is less than 4 Å from one of the cluster iron atoms by a tyrosine residue. Although the spectroscopic results give no indication of tyrosyl cluster ligation, the presence of a tyrosine residue in close proximity to the cluster results in a 38-mV decrease in the midpoint potential of the [4Fe-4S]^2+,+ couple and has a marked effect on the ground state properties of the reduced cluster. The mixed spin [4Fe-4S]⁺ cluster in the wild-type protein, 80% S=3/2 (E/D=0.22, D=+3.3 cm⁻¹) and 20% S=1/2 (g=2.10, 1.87, 1.80), is converted into a homogeneous S=3/2 (E/D=0.30, D=−0.7 cm⁻¹) form in the A33Y variant. As the first example of a pure S=3/2 [4Fe-4S]⁺ cluster in a ferredoxin, this variant affords the opportunity for detailed characterization of the excited electronic properties via VTMCD studies and demonstrates that the protein environment can play a crucial role in determining the ground state properties of [4Fe-4S]⁺ clusters.

【 授权许可】

Unknown