Literature summary for 1.13.11.49 extracted from

de Geus, D.C.; Thomassen, E.A.; Hagedoorn, P.L.; Pannu, N.S.; van Duijn, E.; Abrahamass spectrometry, J.P.
Crystal structure of chlorite dismutase, a detoxifying enzyme producing molecular oxygen (2009), J. Mol. Biol., 387, 192-206.

Search for more literature for 1.13.11.49

Cloned(Commentary)

Cloned (Comment)	Organism
enzyme gene chemically synthesized without signal sequence, cloned into vector pET28a with histidine tag and thrombin cleavage site (removed before crystallization), and overexpressed	Azospira oryzae

Crystallization (Commentary)

Crystallization (Comment)	Organism
thiocyanate inhibited enzyme with incorporated heme, crystals grown from well solution containing 100 mM Mes buffer (pH 5.5), 25% polyethylene glycol monomethyl ether 2000, 0.3 M KSCN, 5% glycerol, 160-260 mM (NH4)2SO4, before data collection crystals are soaked in a solution of the mother liquor including 16% glycerol, crystal is flash-frozen and kept at -173°C (100 K) for multiple anomalous dispersion (MAD) with synchrotron radiation	Azospira oryzae

Crystallization (Comment)

Organism

thiocyanate inhibited enzyme with incorporated heme, crystals grown from well solution containing 100 mM Mes buffer (pH 5.5), 25% polyethylene glycol monomethyl ether 2000, 0.3 M KSCN, 5% glycerol, 160-260 mM (NH4)2SO4, before data collection crystals are soaked in a solution of the mother liquor including 16% glycerol, crystal is flash-frozen and kept at -173°C (100 K) for multiple anomalous dispersion (MAD) with synchrotron radiation

Azospira oryzae

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	activating compound	Azospira oryzae

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
30000	-	4 * 30000 Da, elution position on a gel-filtration column using Stokes radius to determine molecular weight points to globular tetramer, collisional activation of the ions ejects one monomer of the pentamer, so that low-charged tetramer counter complex ions are also observed with native mass spectrometry	Azospira oryzae
30000	-	5 * 30000 Da, tandem mass spectrometry, native mass spectrometry, active state in solution	Azospira oryzae
30000	-	6 * 30000, multiple anomalous dispersion crystal structure, one heme per monomer (histidine 170 axial heme ligand), arginine 183 probably with essential role in substrate positioning and activation, activity of the hexamer not measurable because of the presence of the thiocyanate inhibitor, each monomer appears to be acting on its own, therefore, different quaternary states may be biologically relevant depending on protein concentration, pH, or other factors	Azospira oryzae
30420	-	monomer without heme, native mass spectrometry, enzyme buffer-exchanged to 50 mM ammonium acetate, pH 6.8, with centrifugal filter units with 30 kDa cutoff	Azospira oryzae
31030	-	monomer with heme, native mass spectrometry, enzyme buffer-exchanged to 50 mM ammonium acetate, pH 6.8, with centrifugal filter units with 30 kDa cutoff	Azospira oryzae
31032	-	1 * 30420 without heme, 1 * 31032 with heme	Azospira oryzae
155000	-	pentamer, native mass spectrometry, native mass spectrometry, enzyme buffer-exchanged to 50 mM ammonium acetate, pH 6.8, with centrifugal filter units with 30 kDa cutoff	Azospira oryzae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
chlorite	Azospira oryzae	catalytic mechanism via ferryl species and ClO--anion	chloride + O2	-	?
chlorite	Azospira oryzae GR-1	catalytic mechanism via ferryl species and ClO--anion	chloride + O2	-	?

Organism

Organism	UniProt	Comment	Textmining
Azospira oryzae	E2DI02	-	-
Azospira oryzae GR-1	E2DI02	-	-

Purification (Commentary)

Purification (Comment)	Organism
HiLoad 16/60 Superdex 200 prep-grade column, equilibrated with 20 mM Tris-HCl (pH 7.5) and 135 mM NaCl to determin molecular Stokes radius	Azospira oryzae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
1500	-	100 mM K3PO4 (pH 7.0), 25°C, oxygen removal by bubbling with high-purity argon, measurement with modified Clarke electrode	Azospira oryzae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
chlorite	catalytic mechanism via ferryl species and ClO--anion	Azospira oryzae	chloride + O2	-	?
chlorite	catalytic mechanism via ferryl species and ClO--anion	Azospira oryzae GR-1	chloride + O2	-	?

Subunits

Subunits	Comment	Organism
hexamer	6 * 30000, multiple anomalous dispersion crystal structure, one heme per monomer (histidine 170 axial heme ligand), arginine 183 probably with essential role in substrate positioning and activation, activity of the hexamer not measurable because of the presence of the thiocyanate inhibitor, each monomer appears to be acting on its own, therefore, different quaternary states may be biologically relevant depending on protein concentration, pH, or other factors	Azospira oryzae
monomer	1 * 30420 without heme, 1 * 31032 with heme	Azospira oryzae
pentamer	5 * 30000 Da, tandem mass spectrometry, native mass spectrometry, active state in solution	Azospira oryzae
tetramer	4 * 30000 Da, elution position on a gel-filtration column using Stokes radius to determine molecular weight points to globular tetramer, collisional activation of the ions ejects one monomer of the pentamer, so that low-charged tetramer counter complex ions are also observed with native mass spectrometry	Azospira oryzae

Cofactor

Cofactor	Comment	Organism	Structure
heme	-	Azospira oryzae