A Persulfurated Cysteine Promotes Active Site Reactivity in Azotobacter vinelandii Rhodanese
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Domenico Bordo
Abstract
Active site reactivity and specificity of RhdA, a thiosulfate:cyanide sulfurtransferase (rhodanese) from Azotobacter vinelandii, have been investigated through ligand binding, sitedirected mutagenesis, and Xray crystallographic techniques, in a combined approach. In native RhdA the active site Cys230 is found persulfurated; fluorescence and sulfurtransferase activity measurements show that phosphate anions interact with Cys230 persulfide sulfur atom and modulate activity. Crystallographic analyses confirm that phosphate and hypophosphite anions react with native RhdA, removing the persulfide sulfur atom from the active site pocket. Considering that RhdA and the catalytic subunit of Cdc25 phosphatases share a common threedimensional fold as well as active site Cys (catalytic) and Arg residues, two RhdA mutants carrying a single amino acid insertion at the active site loop were designed and their phosphatase activity tested. The crystallographic and functional results reported here show that specific sulfurtransferase or phosphatase activities are strictly related to precise tailoring of the catalytic loop structure in RhdA and Cdc25 phosphatase, respectively.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
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Artikel in diesem Heft
- Roger D. Kornberg Felix Hoppe-Seyler Lecturer 2001
- The Eukaryotic Gene Transcription Machinery
- Paper of the Year 2000: Award to Eva Estébanez-Perpiñá and Pablo Fuentes-Prior
- GABAC Receptors: A Molecular View
- Recent Advances in Plant MAP Kinase Signalling
- The Chemical Biology of Ras Lipidation
- This Is the End: Processing, Editing and Repair at the tRNA 3-Terminus
- Binding of IRE-BP to Its Cognate RNA Sequence: SFM Studies on a Universal RNA Backbone for the Analysis of RNA-Protein Interaction
- PAF67, a Novel Protein That Is Associated with the Initiation-Competent Form of RNA Polymerase I
- Characterization of RNase P Holoenzymes from Methanococcus jannaschii and Methanothermobacter thermoautotrophicus
- Identification of Functional Regions of Guanylate Cyclase-Activating Protein 1 (GCAP1) Using GCAP1/GCIP Chimeras
- Cooperativity of Binding Epitopes and Receptor Chains in the BMP/TGFß Superfamily
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