Literature summary for 1.14.13.9 extracted from

Mimasu, S.; Yamagishi, H.; Kubo, S.; Kiyohara, M.; Matsuda, T.; Yahata, T.; Thomson, H.A.; Hupp, C.D.; Liu, J.; Okuda, T.; Kakefuda, K.
Full-length in meso structure and mechanism of rat kynurenine 3-monooxygenase inhibition (2021), Commun. Biol., 4, 159 .

Search for more literature for 1.14.13.9

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant enzyme expression in HEK-293 cells	Homo sapiens
sequence comparisons, recombinant expression of GST-tagged full-length KMO (1-478) from baculovirus vector containing a thrombin cleavage site inserted between GST and the N-terminus of KMO, and a TEV site followed by a FLAG tag at the C-terminus with the Bac-to-Bac Baculovirus system in Spodoptera frugiperda (Sf9) cells. Recombinant enzyme expression in HEK-293 cells	Rattus norvegicus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified full-length structure of KMO in its membrane-embedded form complexed with inhibitors 2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid or 4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid, sitting drop vapour diffusion method, mixing of enzyme in lipidic cubic phase formed by mixing a 9:1 monoolein:cholesterol molten lipid mixture and inhibitors, with reservoir solution containing 0.04 M Tris-Cl, pH 7.0, 0.06 M Bis-Tris, pH 6.5, 0.3-0.51 M lithium sulfate, and 34-45% PEG 400 for 2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid and containing 0.04-0.05 M sodium citrate, pH 6.5, or 0.04 M Bis-Tris pH 6.5, 0.05-0.06 M Tris-Cl, pH 7.0, 0.12-0.43 M lithium sulfate, and 34-46% PEG 400 for 4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid, in a 2:3 ratio at 17°C for 1-3 days, X-ray diffraction structure determination and analysis at 3.0 A resolution	Rattus norvegicus
purified full-length structure of KMO in its membrane-embedded form, complexed with 2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid and 4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid at 3.0 A resolution	Homo sapiens

Crystallization (Comment)

Organism

purified full-length structure of KMO in its membrane-embedded form complexed with inhibitors 2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid or 4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid, sitting drop vapour diffusion method, mixing of enzyme in lipidic cubic phase formed by mixing a 9:1 monoolein:cholesterol molten lipid mixture and inhibitors, with reservoir solution containing 0.04 M Tris-Cl, pH 7.0, 0.06 M Bis-Tris, pH 6.5, 0.3-0.51 M lithium sulfate, and 34-45% PEG 400 for 2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid and containing 0.04-0.05 M sodium citrate, pH 6.5, or 0.04 M Bis-Tris pH 6.5, 0.05-0.06 M Tris-Cl, pH 7.0, 0.12-0.43 M lithium sulfate, and 34-46% PEG 400 for 4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid, in a 2:3 ratio at 17°C for 1-3 days, X-ray diffraction structure determination and analysis at 3.0 A resolution

Rattus norvegicus

purified full-length structure of KMO in its membrane-embedded form, complexed with 2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid and 4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid at 3.0 A resolution

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D184A	site-directed mutagenesis, the mutation weakens the beta-sheet dimer interface of the enzyme	Rattus norvegicus
Q187A	site-directed mutagenesis, the mutation weakens the beta-sheet dimer interface of the enzyme	Rattus norvegicus
R380A	site-directed mutagenesis, the mutation has no effect on kynurenine hydroxylation, suggesting that residue R380 does not play a major role in substrate recognition	Rattus norvegicus
Y185P	site-directed mutagenesis, the mutation weakens the beta-sheet dimer interface of the enzyme	Rattus norvegicus

Inhibitors

Inhibitors	Comment	Organism
2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid	-	Homo sapiens
2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid	R380, a residue from the enzyme's C-terminal region, forms hydrogen bonds with the carboxylic acid moiety of the inhibitor, residues R85, Y99 and Y398 also form bonds to 2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid	Rattus norvegicus
3-(4-methyl-5-phenyl-1H-pyrazol-1-yl)benzoic acid	-	Homo sapiens
3-(4-methyl-5-phenyl-1H-pyrazol-1-yl)benzoic acid	-	Rattus norvegicus
3-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid	-	Homo sapiens
3-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid	-	Rattus norvegicus
4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid	-	Homo sapiens
4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid	ligand-binding structure, overview	Rattus norvegicus
6-[3-(4-chloro-3-fluorophenyl)pyridin-2-yl]-1-methylquinazoline-2,4(1H,3H)-dione	-	Homo sapiens
6-[3-(4-chloro-3-fluorophenyl)pyridin-2-yl]-1-methylquinazoline-2,4(1H,3H)-dione	-	Rattus norvegicus
additional information	determinations of inhibition with the purified enzyme and a cell-based assay	Homo sapiens
additional information	determinations of inhibition with the purified enzyme and a cell-based assay, docking studies of KMO representative inhibitors, inhibition mechanism, overview	Rattus norvegicus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
mitochondrial outer membrane	KMO has two transmembrane domains, KMO is actually a single-pass transmembrane protein, with the other transmembrane domain lying laterally along the membrane, where it forms part of the ligand-binding pocket. Membrane-bound structure, overview	Homo sapiens	5741	-
mitochondrial outer membrane	KMO has two transmembrane domains, KMO is actually a single-pass transmembrane protein, with the other transmembrane domain lying laterally along the membrane, where it forms part of the ligand-binding pocket. Membrane-bound structure, overview	Rattus norvegicus	5741	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-kynurenine + NADPH + H+ + O2	Homo sapiens	-	3-hydroxy-L-kynurenine + NADP+ + H2O	-	?
L-kynurenine + NADPH + H+ + O2	Rattus norvegicus	-	3-hydroxy-L-kynurenine + NADP+ + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	O15229	-	-
Rattus norvegicus	O88867	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant FLAG/GST-tagged full-length KMO (1-478) from Spodoptera frugiperda (Sf9) cells by glutathione affinity chromatography, ultrafiltration, and desalting gel filtration, followed by tag cleavage through thrombin, gel filtration, anti-FLAG affinity imuno-chromatography, and again ultrafiltration and gel filtration	Rattus norvegicus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-kynurenine + NADPH + H+ + O2	-	Homo sapiens	3-hydroxy-L-kynurenine + NADP+ + H2O	-	?
L-kynurenine + NADPH + H+ + O2	-	Rattus norvegicus	3-hydroxy-L-kynurenine + NADP+ + H2O	-	?

Subunits

Subunits	Comment	Organism
homodimer	ligand docking and human KMO model construction, overview	Homo sapiens
homodimer	the rat KMO monomer consists of three domains: alpha + beta flavin adenine dinucleotide (FAD)-binding domain I, FPMO enzyme-conserved domain II containing six-stranded beta-sheets, and C-terminal domain III, which is predicted to be comprising two transmembrane domains, although the exact topology remains ambiguous. The C-terminal region is comprising only one transmembrane domain, with the other predicted transmembrane helix lying laterally along the membrane, this helix displays hydrophobic residues on the outer side	Rattus norvegicus

Synonyms

Synonyms	Comment	Organism
Hs-KMO	-	Homo sapiens
KMO	-	Homo sapiens
KMO	-	Rattus norvegicus
Rat-KMO	-	Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens
37	-	assay at	Rattus norvegicus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.9	-	assay at	Homo sapiens
7.9	-	assay at	Rattus norvegicus

Cofactor

Cofactor	Comment	Organism
FAD	-	Homo sapiens
FAD	-	Rattus norvegicus
NADPH	-	Homo sapiens
NADPH	-	Rattus norvegicus

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor
0.00000071	-	pH 7.9, 37°C	Homo sapiens	3-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid
0.0000017	-	pH 7.9, 37°C	Rattus norvegicus	4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid
0.0000038	-	pH 7.9, 37°C	Homo sapiens	2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid
0.0000068	-	pH 7.9, 37°C	Homo sapiens	6-[3-(4-chloro-3-fluorophenyl)pyridin-2-yl]-1-methylquinazoline-2,4(1H,3H)-dione
0.000009	-	pH 7.9, 37°C	Rattus norvegicus	2-(benzyloxy)-5-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid
0.000014	-	pH 7.9, 37°C	Homo sapiens	4-chloro-2-([5-chloro-2-(5-methoxy-1,3-dihydro-2H-isoindol-2-yl)-1,3-thiazole-4-carbonyl](methyl)amino)-5-fluorobenzoic acid
0.00013	-	pH 7.9, 37°C	Rattus norvegicus	6-[3-(4-chloro-3-fluorophenyl)pyridin-2-yl]-1-methylquinazoline-2,4(1H,3H)-dione
0.00015	-	pH 7.9, 37°C	Rattus norvegicus	3-[5-(4-chloro-3-fluorophenyl)-4-methyl-1H-pyrazol-1-yl]benzoic acid
0.00016	-	pH 7.9, 37°C	Homo sapiens	3-(4-methyl-5-phenyl-1H-pyrazol-1-yl)benzoic acid
0.00057	-	pH 7.9, 37°C	Rattus norvegicus	3-(4-methyl-5-phenyl-1H-pyrazol-1-yl)benzoic acid

General Information

General Information	Comment	Organism
malfunction	mutations at the dimeric interface abolish the enzyme activity	Homo sapiens
malfunction	mutations at the dimeric interface abolish the enzyme activity	Rattus norvegicus
additional information	analysis of the extended ligand-binding pocket of in meso KMO and its binding mode	Homo sapiens
additional information	analysis of the extended ligand-binding pocket of in meso KMO and its binding mode	Rattus norvegicus
physiological function	kynurenine 3-monooxygenase (KMO) is a mitochondrial protein involved in the eukaryotic tryptophan catabolic pathway and is linked to various diseases	Homo sapiens
physiological function	kynurenine 3-monooxygenase (KMO) is a mitochondrial protein involved in the eukaryotic tryptophan catabolic pathway and is linked to various diseases	Rattus norvegicus