Literature summary for 1.14.14.21 extracted from

Liu, S.; Zhang, C.; Su, T.; Wei, T.; Zhu, D.; Wang, K.; Huang, Y.; Dong, Y.; Yin, K.; Xu, S.; Xu, P.; Gu, L.
Crystal structure of DszC from Rhodococcus sp. XP at 1.79 A (2014), Proteins, 82, 1708-1720.

Search for more literature for 1.14.14.21

Cloned(Commentary)

Cloned (Comment)	Organism
gene dszC, overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Rhodococcus sp.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type, selenomethionine-labeled, and mutant enzymes, hanging drop vapor diffusion method, mixing of 15-20 mg/ml protein in 10 mM Tris-HCl, pH 8.0, and 100 mM NaCl with an equal volume of reservoir solution containing 0.2 M L2SO4, 0.1 M Bis-Tris, pH 6.5, and 23% w/v PEG 3350, at 20°C, X-ray diffraction structure determination and analysis at 1.79 A resolution, molecular replacement and modelling	Rhodococcus sp.

Crystallization (Comment)

Organism

purified recombinant wild-type, selenomethionine-labeled, and mutant enzymes, hanging drop vapor diffusion method, mixing of 15-20 mg/ml protein in 10 mM Tris-HCl, pH 8.0, and 100 mM NaCl with an equal volume of reservoir solution containing 0.2 M L2SO4, 0.1 M Bis-Tris, pH 6.5, and 23% w/v PEG 3350, at 20°C, X-ray diffraction structure determination and analysis at 1.79 A resolution, molecular replacement and modelling

Rhodococcus sp.

Protein Variants

Protein Variants	Comment	Organism
D392N	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
F161A	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
H388F	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
H391F	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
H92F	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
N129A	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
R282A	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
R370A	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
S163A	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
W205A	site-directed mutagenesis, inactive mutant	Rhodococcus sp.
Y96A	site-directed mutagenesis, inactive mutant	Rhodococcus sp.

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
dibenzothiophene + 2 FMNH2 + 2 O2	Rhodococcus sp.	overall reaction	dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O	-	?
dibenzothiophene + 2 FMNH2 + 2 O2	Rhodococcus sp. XP	overall reaction	dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O	-	?
dibenzothiophene + FMNH2 + O2	Rhodococcus sp.	-	dibenzothiophene-5-oxide + FMN + H2O	-	?
dibenzothiophene + FMNH2 + O2	Rhodococcus sp. XP	-	dibenzothiophene-5-oxide + FMN + H2O	-	?
dibenzothiophene-5-oxide + FMNH2 + O2	Rhodococcus sp.	-	dibenzothiophene-5,5-dioxide + FMN + H2O	-	?
dibenzothiophene-5-oxide + FMNH2 + O2	Rhodococcus sp. XP	-	dibenzothiophene-5,5-dioxide + FMN + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Rhodococcus sp.	Q6WNP1	gene dszC	-
Rhodococcus sp. XP	Q6WNP1	gene dszC	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type, selenomethionine-labeled, and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, and gel filtration	Rhodococcus sp.

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
dibenzothiophene + 2 FMNH2 + 2 O2	overall reaction	Rhodococcus sp.	dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O	-	?
dibenzothiophene + 2 FMNH2 + 2 O2	overall reaction	Rhodococcus sp. XP	dibenzothiophene-5,5-dioxide + 2 FMN + 2 H2O	-	?
dibenzothiophene + FMNH2 + O2	-	Rhodococcus sp.	dibenzothiophene-5-oxide + FMN + H2O	-	?
dibenzothiophene + FMNH2 + O2	-	Rhodococcus sp. XP	dibenzothiophene-5-oxide + FMN + H2O	-	?
dibenzothiophene-5-oxide + FMNH2 + O2	-	Rhodococcus sp.	dibenzothiophene-5,5-dioxide + FMN + H2O	-	?
dibenzothiophene-5-oxide + FMNH2 + O2	-	Rhodococcus sp. XP	dibenzothiophene-5,5-dioxide + FMN + H2O	-	?
additional information	reduced FMN reacts with an oxygen molecule at C4a position of the isoalloxazine ring, producing the C4a-(hydro)peroxyflavin intermediate which is stabilized by residues H391 and S163. H391 may contribute to the formation of the C4a-(hydro)peroxyflavin by acting as a proton donor to the proximal peroxy oxygen, and it might also be involved in the protonation process of the C4a-(hydro)xyflavin, substrate binding and mechanism, molecular docking	Rhodococcus sp.	?	-	?
additional information	reduced FMN reacts with an oxygen molecule at C4a position of the isoalloxazine ring, producing the C4a-(hydro)peroxyflavin intermediate which is stabilized by residues H391 and S163. H391 may contribute to the formation of the C4a-(hydro)peroxyflavin by acting as a proton donor to the proximal peroxy oxygen, and it might also be involved in the protonation process of the C4a-(hydro)xyflavin, substrate binding and mechanism, molecular docking	Rhodococcus sp. XP	?	-	?

Subunits

Subunits	Comment	Organism
homotetramer	enzyme DszC is a tightly associated homotetramer, which can be aptly described as a dimer of dimers	Rhodococcus sp.
More	two distinct conformations occur in the flexible lid loops adjacent to the active site (residue 280-295, between helix alpha9 and alpha10), that are named open and closed state respectively, and might show the status of the free and ligand-bound DszC	Rhodococcus sp.

Synonyms

Synonyms	Comment	Organism
DBT monooxygenase	-	Rhodococcus sp.
dibenzothiophene monooxygenase	-	Rhodococcus sp.
dszC	-	Rhodococcus sp.

Cofactor

Cofactor	Comment	Organism	Structure
FMNH2	reduced FMN reacts with an oxygen molecule at C4a position of the isoalloxazine ring, producing the C4a-(hydro)peroxyflavin intermediate which is stabilized by residues H391 and S163. H391 may contribute to the formation of the C4a-(hydro)peroxyflavin by acting as a proton donor to the proximal peroxy oxygen, and it might also be involved in the protonation process of C4a-(hydro)peroxyflavin, molecular docking simulation and modeling, overview	Rhodococcus sp.

General Information

General Information	Comment	Organism
evolution	DszC with specificity for FMN makes a unique member of the flavin monooxygenase family	Rhodococcus sp.
malfunction	site-directed mutagenesis study shows that mutations in the residues involved either in catalysis or in flavin or substrate-binding result in a complete loss of enzyme activity, suggesting that the accurate positions of flavin and substrate are crucial for the enzyme activity	Rhodococcus sp.
additional information	two distinct conformations occur in the flexible lid loops adjacent to the active site (residue 280-295, between helix alpha9 and alpha10), that are named open and closed state, respectively, and might show the status of the free and ligand-bound DszC	Rhodococcus sp.
physiological function	the dibenzothiophene (DBT) monooxygenase DszC, which is the key initiating enzyme in 4S metabolic pathway, catalyzes sequential sulphoxidation reaction of DBT to DBT sulfoxide (DBTO), then DBT sulfone (DBTO2). Residues H391, and D392 directly participate in catalysis. Residues H92, Y96, N129, F161, S163, W205, R282, R370, and H388 are involved in flavin or substrate-binding	Rhodococcus sp.