Engineering a Three-cysteine, One-histidine Ligand Environment into a New Hyperthermophilic Archaeal Rieske-type [2Fe-2S] Ferredoxin from Sulfolobus solfataricus

Kounosu, Asako; Li, Zhongrui; Cosper, Nathaniel J.; Shokes, Jacob E.; Scott, Robert A.; Imai, Takeo; Urushiyama, Akio; Iwasaki, Toshio

doi:10.1074/jbc.m305923200

Cited by 51 publications

(133 citation statements)

References 74 publications

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“…The 250-cm Ϫ1 band would have a major contribution from symmetric stretching of the two Fe-N (His) bonds, with one or both of the 282 and 296 cm Ϫ1 bands having a significant contribution from asymmetric stretching of the two Fe-N(His) bonds. The available pH dependence and N-isotope shift data for the Rieske-type and mitoNEET proteins argues against the assignment of the band in the 250 -300 cm Ϫ1 region to pure Fe-N(His) stretching mode (42)(43)(44)(45). Rather, Fe-N(His) stretching is distributed over low energy Fe-S stretching modes and internal modes of coordinated cysteine ligands and enhanced via the visible S-to-Fe charge transfer transitions.…”

Section: Rieske-type [2fe-2s] Coordination Occurs In Grx-bola_hmentioning

confidence: 96%

Structural and Spectroscopic Insights into BolA-Glutaredoxin Complexes

Roret

Tsan

Couturier

et al. 2014

Journal of Biological Chemistry

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Background: BolA and glutaredoxin interact together in the iron metabolism. Results: They form two types of heterodimers with different binding surfaces depending on the presence or absence of an iron-sulfur cluster. Conclusion: The function of both proteins is likely modulated by the nature of the interaction. Significance: Understanding the molecular mechanisms responsible for iron sensing is crucial for all iron-mediated cellular processes.

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Section: Rieske-type [2fe-2s] Coordination Occurs In Grx-bola_hmentioning

confidence: 96%

Structural and Spectroscopic Insights into BolA-Glutaredoxin Complexes

Roret

Tsan

Couturier

et al. 2014

Journal of Biological Chemistry

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“…In Rieske-type clusters, Fe1 is coordinated by two cysteines, and Fe2 is coordinated by two histidines (10). In mitoNEET, Fe2 is asymmetrically coordinated by one cysteine and one histidine, which to the best of our knowledge represents a previously unrecognized, naturally occurring coordination pattern for a [2Fe-2S] cluster, although a Rieske-type protein has been successfully engineered to contain three cysteines and one histidine ligands (11). Histidine ligation has been proposed to result in unique redox and spectroscopic properties of Rieske-type clusters and to couple proton and electron transport (12)(13)(14).…”

Section: Structure Of [2fe-2s] Bindingmentioning

confidence: 99%

Crystal structure of human mitoNEET reveals distinct groups of iron–sulfur proteins

Lin

Zhou

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

110

126

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MitoNEET is a protein of unknown function present in the mitochondrial membrane that was recently shown to bind specifically the antidiabetic drug pioglizatone. Here, we report the crystal structure of the soluble domain (residues 32-108) of human mitoNEET at 1.8-Å resolution. The structure reveals an intertwined homodimer, and each subunit was observed to bind a [2Fe-2S] cluster. The [2Fe-2S] ligation pattern of three cysteines and one histidine differs from the known pattern of four cysteines in most cases or two cysteines and two histidines as observed in Rieske proteins. The [2Fe-2S] cluster is packed in a modular structure formed by 17 consecutive residues. The cluster-binding motif is conserved in at least seven distinct groups of proteins from bacteria, archaea, and eukaryotes, which show a consensus sequence of (hb)-C-X1-C-X2-(S/T)-X3-P-(hb)-C-D-X2-H, where hb represents a hydrophobic residue; we term this a CCCH-type [2Fe-2S] binding motif. The nine conserved residues in the motif contribute to iron ligation and structure stabilization. UV-visible absorption spectra indicated that mitoNEET can exist in oxidized and reduced states. Our study suggests an electron transfer function for mitoNEET and for other proteins containing the CCCH motif.2Fe-2S ͉ thiazolidinediones ͉ mitochondria T ype 2 diabetes is a growing global health problem characterized by insulin resistance and pancreatic ␤-cell dysfunction (1). Thiazolidinediones (TZDs) are a class of insulinsensitizing drugs used for treatment of type 2 diabetes (2), which includes rosiglitazone and pioglitazone currently in clinic use. The mechanism of action of TZDs has been generally attributed to their direct activation of peroxisome proliferator-activated receptor-␥, a ligand-binding nuclear receptor important for adipocyte differentiation and glucose homeostasis (3). However, accumulating evidence suggests that TZDs may also exert effects via a peroxisome proliferator-activated receptor-␥-independent pathway, particularly through modulation of mitochondrial activity (4-6). Colca et al. (7) have recently identified a protein in the mitochondrial membrane that crosslinks with photo-affinitylabeled pioglitazone (7). The crosslink could be competed by unlabeled pioglitazone, suggesting specificity in TZD binding. The protein was named mitoNEET because of its mitochondrial location and because of the presence of the sequence motif Asn-Glu-Glu-Thr (''NEET''). MitoNEET has a putative Nterminal transmembrane helix, which likely serves as a membrane anchor, and several invariant cysteine and histidine residues, which suggests that mitoNEET contains a CDGSH-type zinc finger (Fig. 1). However, its sequence is not homologous to any protein or domain of known function. Elucidation of the function and structure of mitoNEET is important to reveal its biological activity, to understand the pharmacology of TZDs, and to aid in the design of more potent antidiabetic drugs. Here, we show by structural characterization that mitoNEET is a previously unrecognized iron-sulfu...

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“…The arf gene coding for the ARF (ORF c06009, DDBJ accession number, AB047031) of S. solfataricus strain P-1 (DSM 1616 T ) has been cloned and sequenced (20). Site-directed mutagenesis was performed by the PCR mutagenesis technique with a QuikChange site-directed mutagenesis kit (Stratagene), using a pET28aARF vector harboring the arf gene (20) as a long template.…”

Section: Methodsmentioning

confidence: 99%

“…Purification of each recombinant holoprotein having a hexahistidinetag at the N terminus was performed as described previously (20,21), except that the heat treatment step (at 65°C for 15-30 min) was omitted for the H44C, H44I/K45C, and double (H44C/H64C) mutants. The recombinant holoprotein was further purified by Sephadex G-75 gel filtration column chromatography (Amersham Biosciences).…”

Section: Methodsmentioning

confidence: 99%

“…Site-directed mutagenesis was performed by the PCR mutagenesis technique with a QuikChange site-directed mutagenesis kit (Stratagene), using a pET28aARF vector harboring the arf gene (20) as a long template. For the double mutant (replacements of His-44 by cysteine and His-64 by cysteine (H44C/H64C)), PCR mutagenesis was carried out in a stepwise manner, with new sets of the following PCR primers: 5Ј-GGT GCG ATT TAT GCG GAT ATG AAT ATA GTC TTG AAA ACG GTG-3Ј and 5Ј-CAC CGT TTT CAA GAC TAT ATT CAT ATC CGC ATA AAT CGC ACC-3Ј for H64C, and 5Ј-GCA TAT TGT CCT TGT AAG GGA AGG AAT CTG G-3Ј and 5Ј-CCA GAT TCC TTC CCT TAC AAG GAC AAT ATG C-3Ј for H44C.…”

Section: Methodsmentioning

confidence: 99%

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Rational Design of a Mononuclear Metal Site into the Archaeal Rieske-type Protein Scaffold

Iwasaki

Kounosu

et al. 2005

Journal of Biological Chemistry

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Proteins containing Rieske-type [2Fe-2S] clusters play essential functions in all three domains of life. We engineered the two histidine ligands to the Rieske-type [2Fe-2S] cluster in the hyperthermophilic archaeal Riesketype ferredoxin from Sulfolobus solfataricus to modify types and spacing of ligands and successfully converted the metal and cluster type at the redox-active site with a minimal structural change to a native Rieske-type protein scaffold. Spectroscopic analyses unambiguously established a rubredoxin-type mononuclear Fe 3؉/2؉ center at the engineered local metal-binding site (Zn 2؉ occupies the iron site depending on the expression conditions). These results show the importance of types and spacing of ligands in the in vivo cluster recognition/insertion/assembly in biological metallosulfur protein scaffolds. We suggest that early ligand substitution and displacement events at the local metal-binding site(s) might have primarily allowed the metal and cluster type conversion in ancestral redox protein modules, which greatly enhanced their capabilities of conducting a wide range of unique redox chemistry in biological electron transfer conduits, using a limited number of basic protein scaffolds.Natural selection allows the utilization of a limited number of protein scaffolds to produce proteins with different types of active sites for various biological catalysis, molecular recognition, and metabolic needs. Metal ions add new functionality to proteins, facilitating some of the most difficult biological catalytic reactions (1-3). For this reason, design and engineering of a specific metalbinding site in natural and de novo protein scaffolds to promote new and specific functionalities are attractive targets in the field of basic and applied protein design research (4, 5).The metallosulfur redox sites, containing sulfurs, usually from cysteinyl side chains, are the most common in biological redox-active metalloproteins (1). In particular, the iron-sulfur (Fe-S) proteins are widely distributed over all living organisms and have been considered to be of early evolutionary origin (2, 6, 7). The mononuclear iron core is the simplest form of Fe-S redox sites, present in small modular proteins such as rubredoxin (Rd) 1 and desulforedoxin. These proteins have the iron atom coordinated in an approximately tetrahedral geometry to the sulfur atoms of four cysteinyl residues (1). Other major forms of protein-bound Fe-S redox sites are polynuclear clusters (such as [2Fe-2S], [3Fe-4S], and [4Fe-4S] clusters) for which some specific synthesis/assembly enzymes are required for in vivo cluster formation and maturation steps (7-10). These sites are also found within complex metalloprotein molecules themselves where they form part of the internal electron transfer conduit to or from the catalytic site (e.g. in some membrane-bound respiratory complexes (11-13)) as a result of modular evolution.Among the biological Fe-S clusters with at least one noncysteinyl ligand, "Rieske-type" [2Fe-2S] clusters are ubiquitous in a vari...

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Engineering a Three-cysteine, One-histidine Ligand Environment into a New Hyperthermophilic Archaeal Rieske-type [2Fe-2S] Ferredoxin from Sulfolobus solfataricus

Cited by 51 publications

References 74 publications

Structural and Spectroscopic Insights into BolA-Glutaredoxin Complexes

Structural and Spectroscopic Insights into BolA-Glutaredoxin Complexes

Crystal structure of human mitoNEET reveals distinct groups of iron–sulfur proteins

Rational Design of a Mononuclear Metal Site into the Archaeal Rieske-type Protein Scaffold

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