Literature summary for 1.14.13.9 extracted from

Gao, J.; Yao, L.; Xia, T.; Liao, X.; Zhu, D.; Xiang, Y.
Biochemistry and structural studies of kynurenine 3-monooxygenase reveal allosteric inhibition by Ro 61-8048 (2017), FASEB J., 32, 2036-2045.

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Application

Application	Comment	Organism
medicine	the enzyme is a potential therapeutic target for neurodegenerative and neurologic disorders	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli and in HEK293T cells	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystals are obtained by hanging-drop vapor diffusion at 20°C. Crystal structures of the complex of the full-length enzyme with the substrate L-kynurenine and in complex with L-kynurenine and Ro 61-8048 at a resolution of 1.85 and 2.34 A, respectively. The crystals of the enzyme-L-kynurenine complex belong to the space group P2(1) with 1 enzyme molecule in the asymmetric unit. The crystal structure of the enzymeL-kynurenine-Ro61-8048 complex belongs to the space group P2(1)2(1)2(1) with 1 pfKMO molecule in the asymmetric unit. The crystal structure of the SeMet enzyme derivative belongs to the space group P2(1) with 2 enzyme molecules in the asymmetric unit	Pseudomonas fluorescens

Crystallization (Comment)

Organism

crystals are obtained by hanging-drop vapor diffusion at 20°C. Crystal structures of the complex of the full-length enzyme with the substrate L-kynurenine and in complex with L-kynurenine and Ro 61-8048 at a resolution of 1.85 and 2.34 A, respectively. The crystals of the enzyme-L-kynurenine complex belong to the space group P2(1) with 1 enzyme molecule in the asymmetric unit. The crystal structure of the enzymeL-kynurenine-Ro61-8048 complex belongs to the space group P2(1)2(1)2(1) with 1 pfKMO molecule in the asymmetric unit. The crystal structure of the SeMet enzyme derivative belongs to the space group P2(1) with 2 enzyme molecules in the asymmetric unit

Pseudomonas fluorescens

Protein Variants

Protein Variants	Comment	Organism
E366Q	2% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
E372A	about 15% of the enzyme activity compared with that of the wild-type enzyme	Pseudomonas fluorescens
E372Q	about 5% of the enzyme activity compared with that of the wild-type enzyme	Pseudomonas fluorescens
M367A	2% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
M367L	13% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
M373A	no activity detected	Pseudomonas fluorescens
M373L	about 60% of the enzyme activity compared with that of the wild-type enzyme	Pseudomonas fluorescens
additional information	the enzyme is systematically truncated according to the sequence alignments and the predicted secondary structures. For the truncations 1-377, 1-379, and 1-394, which have 1 or 2 more predicted alpha-helices than that of the 1-372/374, no or weak protein expression is detected. Although for the truncation 1-430 that has the C-terminal hydrophobic tail removed, no enzymatic activities can be detected but the protein expression is normal. The results indicate that the C-terminal portion is important for both the folding and the enzymatic activity	Homo sapiens
N363A	28% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
N363D	no activity detected	Homo sapiens
N369A	about 65% of the enzyme activity compared with that of the wild-type enzyme	Pseudomonas fluorescens
N369D	no activity detected	Pseudomonas fluorescens
N465A	about 80% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
Q424A	mutation does not greatly affect enzyme activity. Ro 61-8048 shows no inhibition to the pfKMO mutant enzyme	Pseudomonas fluorescens
R84A	no activity detected	Pseudomonas fluorescens
R85A	no activity detected	Homo sapiens
R85K	1% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
Y398A	1% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
Y398F	1% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens
Y404A	no activity detected	Pseudomonas fluorescens
Y404F	about 40% of the enzyme activity compared with that of the wild-type enzyme	Pseudomonas fluorescens
Y98A	no activity detected	Pseudomonas fluorescens
Y98F	about 1% of the enzyme activity compared with that of the wild-type enzyme	Pseudomonas fluorescens
Y99A	no activity detected	Homo sapiens
Y99F	7% of the enzyme activity compared with that of the wild-type enzyme	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism	Structure
Ro 61-8048	allosteric	Homo sapiens
Ro 61-8048	noncompetitive. Theinhibitor is bound in the tunnel at the interface where the N- and C-terminal domains associate	Pseudomonas fluorescens

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
60386	-	calculated from sequence	Homo sapiens
62136	-	native mass spectrum analysis	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	O15229	-	-
Pseudomonas fluorescens	Q84HF5	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	glycosylation has only a marginal effect on enzyme activity	Homo sapiens

Purification (Commentary)

Purification (Comment)	Organism
-	Pseudomonas fluorescens

Substrates and Products (Substrate)

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

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Pseudomonas fluorescens
37	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Pseudomonas fluorescens
8	-	assay at	Homo sapiens

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0135	-	Ro 61-8048	pH and temperature not specified in the publication	Pseudomonas fluorescens

General Information

General Information	Comment	Organism
drug target	the enzyme is a potential therapeutic target for neurodegenerative and neurologic disorders	Homo sapiens