Structural rearrangements of the two domains of Azotobacter vinelandii rhodanese upon sulfane sulfur release: essential molecular dynamics, NMR relaxation and deuterium exchange on the uniformly labeled protein

Cicero, Daniel O.; Melino, Sonia; Orsale, Maria Vittoria; Brancato, Giuseppe; Amadei, Andrea; Forlani, Fabio; Pagani, Silvia; Paci, Maurizio

doi:10.1016/j.ijbiomac.2003.08.010

Cited by 6 publications

(4 citation statements)

References 38 publications

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“…These results are different from the previous relaxation study on A. vinelandii rhodanese, in which the sulfur-free and sulfur-loaded forms showed similar relaxation rates .…”

Section: Resultscontrasting

confidence: 99%

“…The unanalyzed residues included four proline residues that have no amide protons, the unassigned residues, and eight residues that were either overlapped or too weak to be analyzed. The experimentally determined R 1 , R 2 , and { 1 H}- 15 Vinelandii rhodanese, in which the sulfur-free and sulfurloaded forms showed similar relaxation rates (48). Rotational Diffusion Anisotropy.…”

Section: Chemical-shift Assignments and Characterization Of Pspementioning

confidence: 87%

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Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese^,

Li¹,

Yang

Xue

et al. 2008

Biochemistry

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Rhodanese catalyzes the sulfur-transfer reaction that transfers sulfur from thiosulfate to cyanide by a double-displacement mechanism, in which an active cysteine residue plays a central role. Previous studies indicated that the phage-shock protein E (PspE) from Escherichia coli is a rhodanese composed of a single active domain and is the only accessible rhodanese among the three single-domain rhodaneses in E. coli. To understand the catalytic mechanism of rhodanese at the molecular level, we determined the solution structures of the sulfur-free and persulfide-intermediate forms of PspE by nuclear magnetic resonance (NMR) spectroscopy and identified the active site by NMR titration experiments. To obtain further insights into the catalytic mechanism, we studied backbone dynamics by NMR relaxation experiments. Our results demonstrated that the overall structures in both sulfur-free and persulfide-intermediate forms are highly similar, suggesting that no significant conformational changes occurred during the catalytic reaction. However, the backbone dynamics revealed that the motional properties of PspE in its sulfur-free form are different from the persulfide-intermediate state. The conformational exchanges are largely enhanced in the persulfide-intermediate form of PspE, especially around the active site. The present structural and biochemical studies in combination with backbone dynamics provide further insights in understanding the catalytic mechanism of rhodanese.

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“…These results are different from the previous relaxation study on A. vinelandii rhodanese, in which the sulfur-free and sulfur-loaded forms showed similar relaxation rates .…”

Section: Resultscontrasting

confidence: 99%

Section: Chemical-shift Assignments and Characterization Of Pspementioning

confidence: 87%

Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese^,

Li¹,

Yang

Xue

et al. 2008

Biochemistry

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“…However, the in vivo role of rhodanese has not been well established. Notably, the structure of sulfur-free rhodanese and its persulfide-containing form have been solved for a number of organisms [34,36]. A comparison of the two enzyme species indicates a significant conformational rearrangement in the vicinity of the catalytic cysteine upon persulfide formation.…”

Section: Discussionmentioning

confidence: 99%

A Conserved Mechanism for Sulfonucleotide Reduction

et al. 2005

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Sulfonucleotide reductases are a diverse family of enzymes that catalyze the first committed step of reductive sulfur assimilation. In this reaction, activated sulfate in the context of adenosine-5′-phosphosulfate (APS) or 3′-phosphoadenosine 5′-phosphosulfate (PAPS) is converted to sulfite with reducing equivalents from thioredoxin. The sulfite generated in this reaction is utilized in bacteria and plants for the eventual production of essential biomolecules such as cysteine and coenzyme A. Humans do not possess a homologous metabolic pathway, and thus, these enzymes represent attractive targets for therapeutic intervention. Here we studied the mechanism of sulfonucleotide reduction by APS reductase from the human pathogen Mycobacterium tuberculosis, using a combination of mass spectrometry and biochemical approaches. The results support the hypothesis of a two-step mechanism in which the sulfonucleotide first undergoes rapid nucleophilic attack to form an enzyme-thiosulfonate (E-Cys-S-SO3 −) intermediate. Sulfite is then released in a thioredoxin-dependent manner. Other sulfonucleotide reductases from structurally divergent subclasses appear to use the same mechanism, suggesting that this family of enzymes has evolved from a common ancestor.

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“…Water proton NMR relaxation studies [ 53 ] and 35 Cl NMR relaxation studies [ 54 ] on eukaryotic Rhobov report that there are significant changes in the exposure to solvent or to anion binding for the two catalytic states ES and E. These results have been interpreted as due to important interdomain reorientation(s) between the two structural domains of the enzyme upon the catalytic cycle [ 54 ]. However, 15 N NMR relaxation studies together with essential dynamics studies on the prokaryotic TST from Azotobacter vinelandii (RhdA) [ 55 , 56 ] did not show large differences between the two forms indicating that only small conformational rearrangements, probably confined around the active site, occur between the ES and E form. However, all the structural studies on both eukaryotic and prokaryotic TST proteins with double domains are in agreement with an enhanced solvent accessibility in the E form.…”

Section: Structure and Function Of Tstmentioning

confidence: 99%

Thiosulfate-Cyanide Sulfurtransferase a Mitochondrial Essential Enzyme: From Cell Metabolism to the Biotechnological Applications

Buonvino

Arciero

Melino

2022

IJMS

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Thiosulfate: cyanide sulfurtransferase (TST), also named rhodanese, is an enzyme widely distributed in both prokaryotes and eukaryotes, where it plays a relevant role in mitochondrial function. TST enzyme is involved in several biochemical processes such as: cyanide detoxification, the transport of sulfur and selenium in biologically available forms, the restoration of iron–sulfur clusters, redox system maintenance and the mitochondrial import of 5S rRNA. Recently, the relevance of TST in metabolic diseases, such as diabetes, has been highlighted, opening the way for research on important aspects of sulfur metabolism in diabetes. This review underlines the structural and functional characteristics of TST, describing the physiological role and biomedical and biotechnological applications of this essential enzyme.

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Structural rearrangements of the two domains of Azotobacter vinelandii rhodanese upon sulfane sulfur release: essential molecular dynamics, NMR relaxation and deuterium exchange on the uniformly labeled protein

Cited by 6 publications

References 38 publications

Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese^,

Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese^,

A Conserved Mechanism for Sulfonucleotide Reduction

Thiosulfate-Cyanide Sulfurtransferase a Mitochondrial Essential Enzyme: From Cell Metabolism to the Biotechnological Applications

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Structural rearrangements of the two domains of Azotobacter vinelandii rhodanese upon sulfane sulfur release: essential molecular dynamics, NMR relaxation and deuterium exchange on the uniformly labeled protein

Cited by 6 publications

References 38 publications

Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese,

Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese,

A Conserved Mechanism for Sulfonucleotide Reduction

Thiosulfate-Cyanide Sulfurtransferase a Mitochondrial Essential Enzyme: From Cell Metabolism to the Biotechnological Applications

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Product

Resources

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Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese^,

Solution Structures and Backbone Dynamics of Escherichia coli Rhodanese PspE in Its Sulfur-Free and Persulfide-Intermediate Forms: Implications for the Catalytic Mechanism of Rhodanese^,