Identification of persulfide-binding and disulfide-forming cysteine residues in the NifS-like domain of the molybdenum cofactor sulfurase ABA3 by cysteine-scanning mutagenesis.

2.50
HDL Handle:
http://hdl.handle.net/10033/245672
Title:
Identification of persulfide-binding and disulfide-forming cysteine residues in the NifS-like domain of the molybdenum cofactor sulfurase ABA3 by cysteine-scanning mutagenesis.
Authors:
Lehrke, Markus; Rump, Steffen; Heidenreich, Torsten; Wissing, Josef; Mendel, Ralf R; Bittner, Florian
Abstract:
The Moco (molybdenum cofactor) sulfurase ABA3 from Arabidopsis thaliana catalyses the sulfuration of the Moco of aldehyde oxidase and xanthine oxidoreductase, which represents the final activation step of these enzymes. ABA3 consists of an N-terminal NifS-like domain that exhibits L-cysteine desulfurase activity and a C-terminal domain that binds sulfurated Moco. The strictly conserved Cys430 in the NifS-like domain binds a persulfide intermediate, which is abstracted from the substrate L-cysteine and finally needs to be transferred to the Moco of aldehyde oxidase and xanthine oxidoreductase. In addition to Cys⁴³⁰, another eight cysteine residues are located in the NifS-like domain, with two of them being highly conserved among Moco sulfurase proteins and, at the same time, being in close proximity to Cys⁴³⁰. By determination of the number of surface-exposed cysteine residues and the number of persulfide-binding cysteine residues in combination with the sequential substitution of each of the nine cysteine residues, a second persulfide-binding cysteine residue, Cys²⁰⁶, was identified. Furthermore, the active-site Cys⁴³⁰ was found to be located on top of a loop structure, formed by the two flanking residues Cys⁴²⁸ and Cys⁴³⁵, which are likely to form an intramolecular disulfide bridge. These findings are confirmed by a structural model of the NifS-like domain, which indicates that Cys⁴²⁸ and Cys⁴³⁵ are within disulfide bond distance and that a persulfide transfer from Cys⁴³⁰ to Cys²⁰⁶ is indeed possible.
Affiliation:
Department of Plant Biology, Braunschweig University of Technology, Humboldtstrasse 1, 38023 Braunschweig, Germany.
Citation:
Identification of persulfide-binding and disulfide-forming cysteine residues in the NifS-like domain of the molybdenum cofactor sulfurase ABA3 by cysteine-scanning mutagenesis. 2012, 441 (3):823-32 Biochem. J.
Journal:
The Biochemical journal
Issue Date:
1-Feb-2012
URI:
http://hdl.handle.net/10033/245672
DOI:
10.1042/BJ20111170
PubMed ID:
22004669
Type:
Article
Language:
en
ISSN:
1470-8728
Appears in Collections:
Publications of RG Cellular Proteome Research (CPRO)

Full metadata record

DC FieldValueLanguage
dc.contributor.authorLehrke, Markusen_GB
dc.contributor.authorRump, Steffenen_GB
dc.contributor.authorHeidenreich, Torstenen_GB
dc.contributor.authorWissing, Josefen_GB
dc.contributor.authorMendel, Ralf Ren_GB
dc.contributor.authorBittner, Florianen_GB
dc.date.accessioned2012-09-24T14:38:33Z-
dc.date.available2012-09-24T14:38:33Z-
dc.date.issued2012-02-01-
dc.identifier.citationIdentification of persulfide-binding and disulfide-forming cysteine residues in the NifS-like domain of the molybdenum cofactor sulfurase ABA3 by cysteine-scanning mutagenesis. 2012, 441 (3):823-32 Biochem. J.en_GB
dc.identifier.issn1470-8728-
dc.identifier.pmid22004669-
dc.identifier.doi10.1042/BJ20111170-
dc.identifier.urihttp://hdl.handle.net/10033/245672-
dc.description.abstractThe Moco (molybdenum cofactor) sulfurase ABA3 from Arabidopsis thaliana catalyses the sulfuration of the Moco of aldehyde oxidase and xanthine oxidoreductase, which represents the final activation step of these enzymes. ABA3 consists of an N-terminal NifS-like domain that exhibits L-cysteine desulfurase activity and a C-terminal domain that binds sulfurated Moco. The strictly conserved Cys430 in the NifS-like domain binds a persulfide intermediate, which is abstracted from the substrate L-cysteine and finally needs to be transferred to the Moco of aldehyde oxidase and xanthine oxidoreductase. In addition to Cys⁴³⁰, another eight cysteine residues are located in the NifS-like domain, with two of them being highly conserved among Moco sulfurase proteins and, at the same time, being in close proximity to Cys⁴³⁰. By determination of the number of surface-exposed cysteine residues and the number of persulfide-binding cysteine residues in combination with the sequential substitution of each of the nine cysteine residues, a second persulfide-binding cysteine residue, Cys²⁰⁶, was identified. Furthermore, the active-site Cys⁴³⁰ was found to be located on top of a loop structure, formed by the two flanking residues Cys⁴²⁸ and Cys⁴³⁵, which are likely to form an intramolecular disulfide bridge. These findings are confirmed by a structural model of the NifS-like domain, which indicates that Cys⁴²⁸ and Cys⁴³⁵ are within disulfide bond distance and that a persulfide transfer from Cys⁴³⁰ to Cys²⁰⁶ is indeed possible.en_GB
dc.language.isoenen
dc.rightsArchived with thanks to The Biochemical journalen_GB
dc.subject.meshArabidopsisen_GB
dc.subject.meshArabidopsis Proteinsen_GB
dc.subject.meshBacterial Proteinsen_GB
dc.subject.meshCatalytic Domainen_GB
dc.subject.meshCoenzymesen_GB
dc.subject.meshCysteineen_GB
dc.subject.meshDisulfidesen_GB
dc.subject.meshHumansen_GB
dc.subject.meshMetalloproteinsen_GB
dc.subject.meshModels, Biologicalen_GB
dc.subject.meshModels, Molecularen_GB
dc.subject.meshMutagenesis, Site-Directeden_GB
dc.subject.meshProtein Bindingen_GB
dc.subject.meshProtein Interaction Domains and Motifsen_GB
dc.subject.meshProtein Interaction Mapsen_GB
dc.subject.meshPteridinesen_GB
dc.subject.meshSequence Homologyen_GB
dc.subject.meshStructure-Activity Relationshipen_GB
dc.subject.meshSulfidesen_GB
dc.subject.meshSulfurtransferasesen_GB
dc.titleIdentification of persulfide-binding and disulfide-forming cysteine residues in the NifS-like domain of the molybdenum cofactor sulfurase ABA3 by cysteine-scanning mutagenesis.en
dc.typeArticleen
dc.contributor.departmentDepartment of Plant Biology, Braunschweig University of Technology, Humboldtstrasse 1, 38023 Braunschweig, Germany.en_GB
dc.identifier.journalThe Biochemical journalen_GB
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