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Literature summary extracted from

  • Shen, J.; Keithly, M.E.; Armstrong, R.N.; Higgins, K.A.; Edmonds, K.A.; Giedroc, D.P.
    Staphylococcus aureus CstB is a novel multidomain persulfide dioxygenase-sulfurtransferase involved in hydrogen sulfide detoxification (2015), Biochemistry, 54, 4542-4554 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.11.18 gene cst, CstB is encoded by the cst operon and is under the transcriptional control of the persulfide sensor CstR and H2S, recombinant expression of full-length wild-type and mutant enzymes and of the N-terminal PDO-RHD domain in Escherichia coli strain Rosetta (DE3) Staphylococcus aureus

Protein Variants

EC Number Protein Variants Comment Organism
1.13.11.18 C201S site-directed mutagenesis, the mutant protein structure is similar to the wild-type, but it shows reduced activity Staphylococcus aureus
1.13.11.18 C201S/C408S site-directed mutagenesis Staphylococcus aureus
1.13.11.18 C2S site-directed mutagenesis, the mutant is larger than the wild-type and is a hexamer Staphylococcus aureus
1.13.11.18 C408S site-directed mutagenesis, the mutant is larger than the wild-type and is a hexamer. Substitution of the presumed Rhod active-site C408 also appears to destabilize the native protein fold. Catalytically inactive mutant Staphylococcus aureus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.11.18 0.0011
-
 CoASSH recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 0.0018
-
 bacillithiol persulfide recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 0.0078
-
 S-sulfanylglutathione recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 0.02
-
 S-sulfanyl-L-cysteine recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.13.11.18 Fe2+ required Staphylococcus aureus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.13.11.18 226000
-
 recombinant full-length Fe(II)-loaded wild-type CstB, gel filtration Staphylococcus aureus
1.13.11.18 298000
-
 recombinant full-length Fe(II)-loaded CstB mutant C408S, gel filtration Staphylococcus aureus

Organism

EC Number Organism UniProt Comment Textmining
1.13.11.18 Staphylococcus aureus G5ELA4
-
-

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.13.11.18 recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and cleavage of the His-tag by TEV protease, followed by another step of nickel affinity chromatography, and gel filtration to over 95% purity Staphylococcus aureus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.13.11.18 bacillithiol persulfide + O2
-
 Staphylococcus aureus bacillithiol + sulfite
-
 ?
1.13.11.18 CoASSH + O2
-
 Staphylococcus aureus CoA + sulfite
-
 ?
1.13.11.18 additional information coupled persulfide dioxygenase-persulfide transferase (cPDO-PT) activity assay producing thiosulfate as product preferentially from CoASSH and bacillithiol persulfide. CstB does not catalyze the formation of thiosulfate from reduced low molecular weight thiol, oxidized low molecular weight disulfide or Na2S. Cysteine persulfide (CSSH) and glutathione persulfide (GSSH) also serve as persulfide substrates for wild-type CstB but are poorer substrates Staphylococcus aureus ?
-
 ?
1.13.11.18 S-sulfanyl-L-cysteine + O2
-
 Staphylococcus aureus L-cysteine + sulfite
-
 ?
1.13.11.18 S-sulfanylglutathione + O2
-
 Staphylococcus aureus glutathione + sulfite
-
 ?

Subunits

EC Number Subunits Comment Organism
1.13.11.18 More enzyme CstB is predicted to be a three-domain enzyme containing an N-terminal metallo-beta-lactamase-like (MBL), non-heme Fe(II)-containing PDO domain, followed by a pseudo-rhodanese homology (RHD) domain, and a conventional C-terminal rhodanese (Rhod) domain Staphylococcus aureus
1.13.11.18 tetramer 4 * 49276, wild-type enzyme, mass spectrometry, 4 * 49277, sequence calculation Staphylococcus aureus

Synonyms

EC Number Synonyms Comment Organism
1.13.11.18 cPDO-PT
-
 Staphylococcus aureus
1.13.11.18 CstB
-
 Staphylococcus aureus
1.13.11.18 Fe(II)-containing persulfide dioxygenase
-
 Staphylococcus aureus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.13.11.18 25
-
 coupled persulfide dioxygenase-persulfide transferase activity assay at Staphylococcus aureus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.13.11.18 2 8 S-sulfanyl-L-cysteine recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 6.5
-
 S-sulfanylglutathione recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 19.3
-
 CoASSH recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 19.7
-
 bacillithiol persulfide recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.11.18 6
-
 coupled persulfide dioxygenase-persulfide transferase activity assay at Staphylococcus aureus

Expression

EC Number Organism Comment Expression
1.13.11.18 Staphylococcus aureus gene cstB is encoded in the cst operon and is under the transcriptional control of the persulfide sensor CstR and H2S additional information

General Information

EC Number General Information Comment Organism
1.13.11.18 evolution CstB is a multifunctional Fe(II)-containing persulfide dioxygenase (PDO), analogous to the vertebrate protein ETHE1, ethylmalonic encephalopathy protein 1 Staphylococcus aureus
1.13.11.18 malfunction stB C201S and C408S mutants are unable to rescue a NaHS-induced DELTAcstB Staphylococcus aureus
1.13.11.18 physiological function the enzyme is a multidomain persulfide dioxygenase-sulfurtransferase, a multifunctional Fe(II)-containing persulfide dioxygenase. Enzyme CstB oxidizes major cellular low molecular weight persulfide substrates from Staphylococcus aureus, coenzyme A persulfide (CoASSH) and bacillithiol persulfide (BSSH), directly to generate thiosulfate and reduced thiols, thereby avoiding the cellular toxicity of sulfite. Both residue Cys201 in the N-terminal PDO domain (CstBPDO) and residue Cys408 in the C-terminal rhodanese domain (CstBRhod) strongly enhance the thiosulfate generating activity of CstB. CstB also possesses persulfide transferase (reverse rhodanese) activity which generates thiosulfate when provided with LMW persulfides and sulfite, as well as conventional thiosulfate transferase (TST, rhodanese) activity. Both activities require residue Cys408. CstB protects Staphylococcus aureus against H2S toxicity Staphylococcus aureus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.13.11.18 833
-
 S-sulfanylglutathione recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 1400
-
 S-sulfanyl-L-cysteine recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 10944
-
 bacillithiol persulfide recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus
1.13.11.18 17545
-
 CoASSH recombinant wild-type enzyme CstB, pH 6.0, 25°C, coupled persulfide dioxygenase-persulfide transferase activity Staphylococcus aureus