Biotin synthase mechanism: on the origin of sulphur

Bui, Bernadette Tse Sum; Florentin, D.; Fournier, Françoise; Ploux, Olivier; Méjean, Annick; Marquet, Andrée

doi:10.1016/s0014-5793(98)01464-1

Cited by 111 publications

(115 citation statements)

References 40 publications

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“…Preparation of Apo and Holo Wild-type Biotin Synthase and Mutants-This was carried out using methods previously described (14,20).…”

Section: Methodsmentioning

confidence: 99%

“…ICP-AES analysis and UV-visible spectroscopy confirmed the absence of iron in the proteins. The samples were then reconstituted by incubation with FeCl 3 and Na 2 S under similar conditions to those described previously (14,20).…”

Section: Preparation Of Wild-type and His 6 -Tagged Apoenzymes And Thmentioning

confidence: 99%

“…sure (13). Recent studies indicate that a sulfur atom of the [Fe-S] cluster of biotin synthase, rather than cysteine itself, may act as the actual sulfur donor, suggesting that the [Fe-S] cluster of the enzyme must be regenerated prior to the next reaction (14). Thus biotin synthase could be regarded as both a reagent and a catalyst.…”

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confidence: 99%

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Identification of the [Fe-S] Cluster-binding Residues of Escherichia coli Biotin Synthase

McIver

Baxter

Campopiano

2000

Journal of Biological Chemistry

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The gene encoding Escherichia coli biotin synthase (bioB) has been expressed as a histidine fusion protein, and the protein was purified in a single step using immobilized metal affinity chromatography. The His 6 -tagged protein was fully functional in in vitro and in vivo biotin production assays. Analysis of all the published bioB sequences identified a number of conserved residues. Single point mutations, to either serine or threonine, were carried out on the four conserved (Cys-53, Cys-57, Cys-60, and Cys-188) and one non-conserved (Cys-288) cysteine residues, and the purified mutant proteins were tested both for ability to reconstitute the [2Fe-2S] clusters of the native (oxidized) dimer and enzymatic activity. The C188S mutant was insoluble. The wild-type and four of the mutant proteins were characterized by UV-visible spectroscopy, metal and sulfide analysis, and both in vitro and in vivo biotin production assays. The molecular masses of all proteins were verified using electrospray mass spectrometry. The results indicate that the His 6 tag and the C288T mutation have no effect on the activity of biotin synthase when compared with the wild-type protein. The C53S, C57S, and C60S mutant proteins, both as prepared and reconstituted, were unable to covert dethiobiotin to biotin in vitro and in vivo. We conclude that three of the conserved cysteine residues (Cys-53, Cys-57, and Cys-60), all of which lie in the highly conserved "cysteine box" motif, are crucial for [Fe-S] cluster binding, whereas Cys-188 plays a hitherto unknown structural role in biotin synthase.

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“…Preparation of Apo and Holo Wild-type Biotin Synthase and Mutants-This was carried out using methods previously described (14,20).…”

Section: Methodsmentioning

confidence: 99%

Section: Preparation Of Wild-type and His 6 -Tagged Apoenzymes And Thmentioning

confidence: 99%

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Identification of the [Fe-S] Cluster-binding Residues of Escherichia coli Biotin Synthase

McIver

Baxter

Campopiano

2000

Journal of Biological Chemistry

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“…A [2Fe-2S] 2+ cluster is bound in close proximity to dethiobiotin within the core of the (αβ) 8 barrel fold. Both isotope labeling and spectroscopic data suggest that a sulfide from this cluster becomes incorporated into dethiobiotin [9,13,14], generating the thiophane ring of biotin. However, the cluster is more than just a binding site for sulfide.…”

Section: Discussionmentioning

confidence: 99%

“…BioB expressed in media containing 35 S-cysteine [11], or apoBioB reconstituted with Fe 3+ and 34 S-sulfide or selenide [12][13][14], results in an enzyme with an isotope-or selenide-labeled [2Fe-2S] 2+ cluster whose turnover produces biotin that incorporates the heavy-atom label in 60-80% yield. When turnover of BioB is monitored by UV/visible spectroscopy, the absorption band at ∼450 nm associated with the [2Fe-2S] 2+ cluster disappears at a rate that is similar to the rate of biotin formation [9].…”

Section: Introductionmentioning

confidence: 99%

Loss of iron–sulfur clusters from biotin synthase as a result of catalysis promotes unfolding and degradation

Reyda

Dippold

Dotson

et al. 2008

Archives of Biochemistry and Biophysics

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Biotin synthase (BioB) is an S-adenosylmethionine radical enzyme that catalyzes addition of sulfur to dethiobiotin to form the biotin thiophane ring. In vitro, E. coli BioB is active for only one turnover, † This research has been supported by the NIH (R01 GM059175 to J.T.J.) and a fellowship from the David and Lucille Packard Foundation (J.T.J. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.Supporting Information Available MALDI mass spectra of tryptic fragments derived from limited proteolysis of apoBioB and 2Fe-BioB.

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Iron–Sulfur Proteins

Johnson

Smith

2005

Encyclopedia of Inorganic Chemistry

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Iron–sulfur proteins contain iron coordinated by at least one sulfur ligand and include proteins with mononuclear Fe centers with partial or complete cysteinyl sulfur ligation as well as proteins containing clusters of iron and inorganic sulfide. This review summarizes the structural, electronic, and redox properties of protein‐bound mononuclear FeS centers and FeS clusters containing [2Fe2S], [3Fe4S], and [4Fe4S] cores. In addition to the ubiquitous role in mediating biological electron transport, the roles of FeS centers have proliferated to include coupling of electron and proton transfer, substrate binding and activation, determining protein structure, regulation of gene expression and enzymatic activity, disulfide reduction, and iron, electron, or cluster storage. The diverse roles of FeS clusters are illustrated by discussion of specific systems: the mitochondrial and photosynthetic electron transport chains and a wide range of soluble redox enzymes and proteins; the active sites of nitrile hydratase, aconitase‐type (de)hydratases, superoxide reductase (SOR), radical‐SAM (S‐adenosylmethionine) enzymes, NiFe‐ and Fe‐hydrogenase, sulfite and nitrite reductase, nitrogenase, CO dehydrogenase, and acetyl‐CoA synthase; the structural FeS centers in DNA repair enzymes; the regulatory roles of FeS centers in the SoxR, fumarate–nitrate reduction (FNR), IscR/SufR, and iron‐regulatory protein (IRP) proteins and in selected enzymes; the two classes of FeS cluster containing disulfide reductases typified by ferredoxin–thioredoxin reductase (FTR) and heterodisulfide reductase (HDR); and the role of 8Fe ferredoxins and polyferredoxins in iron, electron, or cluster storage.

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Biotin synthase mechanism: on the origin of sulphur

Cited by 111 publications

References 40 publications

Identification of the [Fe-S] Cluster-binding Residues of Escherichia coli Biotin Synthase

Identification of the [Fe-S] Cluster-binding Residues of Escherichia coli Biotin Synthase

Loss of iron–sulfur clusters from biotin synthase as a result of catalysis promotes unfolding and degradation

Iron–Sulfur Proteins

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