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- Title
Mutational Analysis of the Cysteine-Rich Region of the Iron-Responsive GATA Factor Fep1. Role of Individual Cysteines as [2Fe-2S] Cluster Ligands.
- Authors
Patti, Maria Carmela Bonaccorsi di; Cutone, Antimo; Musci, Giovanni
- Abstract
Fep1, the iron-dependent GATA-type transcriptional repressor of the methylotrophic yeast Pichia pastoris, has a dimeric structure and binds an iron-sulfur cluster of the [2Fe-2S] type. In this work, we extend the characterization of this protein by analysis of the optical and CD spectroscopic properties of a set of mutants where cysteines within the conserved Cys-X5-Cys-X8-Cys-X2-Cys motif have been targeted, in order to evaluate their role as [2Fe-2S] ligands. The results suggest that all four cysteine residues are essential because replacing them with serines in different combinations invariably produces a protein unable to correctly bind the [2Fe-2S] cluster.
- Subjects
CYSTEINE; GATA proteins; PICHIA pastoris; IRON-sulfur proteins; CIRCULAR dichroism
- Publication
Cell Biochemistry & Biophysics, 2018, Vol 76, Issue 3, p339
- ISSN
1085-9195
- Publication type
Article
- DOI
10.1007/s12013-018-0842-9