Dual role of the carboxyl-terminal region of pig liver l-kynurenine 3-monooxygenase: Mitochondrial-targeting signal and enzymatic activity

Kumiko Hirai, Hidehito Kuroyanagi, Yoshitaka Tatebayashi, Yoshitaka Hayashi, Kanako Hirabayashi-Takahashi, Kuniaki Saito, Seiich Haga, Tomihiko Uemura, Susumu Izumi

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

l-kynurenine 3-monooxygenase (KMO) is an NAD(P)H-dependent flavin monooxygenase that catalyses the hydroxylation of l-kynurenine to 3-hydroxykynurenine, and is localized as an oligomer in the mitochondrial outer membrane. In the human brain, KMO may play an important role in the formation of two neurotoxins, 3-hydroxykynurenine and quinolinic acid, both of which provoke severe neurodegenerative diseases. In mosquitos, it plays a role in the formation both of eye pigment and of an exflagellation-inducing factor (xanthurenic acid). Here, we present evidence that the C-terminal region of pig liver KMO plays a dual role. First, it is required for the enzymatic activity. Second, it functions as a mitochondrial targeting signal as seen in monoamine oxidase B (MAO B) or outer membrane cytochrome b5. The first role was shown by the comparison of the enzymatic activity of two mutants (C-terminally FLAG-tagged KMO and carboxyl-terminal truncation form, KMOΔC50) with that of the wild-type enzyme expressed in COS-7 cells. The second role was demonstrated with fluorescence microscopy by the comparison of the intracellular localization of the wild-type, three carboxyl-terminal truncated forms (ΔC20, ΔC30 and ΔC50), C-terminally FLAG-tagged wild-type and a mutant KMO, where two arginine residues, Arg461-Arg462, were replaced with Ser residues.

Original languageEnglish
Pages (from-to)639-650
Number of pages12
JournalJournal of Biochemistry
Volume148
Issue number6
DOIs
Publication statusPublished - 01-12-2010
Externally publishedYes

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Kynurenine 3-Monooxygenase
Liver
Swine
Neurodegenerative diseases
Quinolinic Acid
Kynurenine
Membranes
Cytochromes b5
Hydroxylation
Fluorescence microscopy
COS Cells
Monoamine Oxidase
Neurotoxins
Mitochondrial Membranes
Mixed Function Oxygenases
Culicidae
Fluorescence Microscopy
Oligomers
Pigments
Neurodegenerative Diseases

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Hirai, Kumiko ; Kuroyanagi, Hidehito ; Tatebayashi, Yoshitaka ; Hayashi, Yoshitaka ; Hirabayashi-Takahashi, Kanako ; Saito, Kuniaki ; Haga, Seiich ; Uemura, Tomihiko ; Izumi, Susumu. / Dual role of the carboxyl-terminal region of pig liver l-kynurenine 3-monooxygenase : Mitochondrial-targeting signal and enzymatic activity. In: Journal of Biochemistry. 2010 ; Vol. 148, No. 6. pp. 639-650.
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abstract = "l-kynurenine 3-monooxygenase (KMO) is an NAD(P)H-dependent flavin monooxygenase that catalyses the hydroxylation of l-kynurenine to 3-hydroxykynurenine, and is localized as an oligomer in the mitochondrial outer membrane. In the human brain, KMO may play an important role in the formation of two neurotoxins, 3-hydroxykynurenine and quinolinic acid, both of which provoke severe neurodegenerative diseases. In mosquitos, it plays a role in the formation both of eye pigment and of an exflagellation-inducing factor (xanthurenic acid). Here, we present evidence that the C-terminal region of pig liver KMO plays a dual role. First, it is required for the enzymatic activity. Second, it functions as a mitochondrial targeting signal as seen in monoamine oxidase B (MAO B) or outer membrane cytochrome b5. The first role was shown by the comparison of the enzymatic activity of two mutants (C-terminally FLAG-tagged KMO and carboxyl-terminal truncation form, KMOΔC50) with that of the wild-type enzyme expressed in COS-7 cells. The second role was demonstrated with fluorescence microscopy by the comparison of the intracellular localization of the wild-type, three carboxyl-terminal truncated forms (ΔC20, ΔC30 and ΔC50), C-terminally FLAG-tagged wild-type and a mutant KMO, where two arginine residues, Arg461-Arg462, were replaced with Ser residues.",
author = "Kumiko Hirai and Hidehito Kuroyanagi and Yoshitaka Tatebayashi and Yoshitaka Hayashi and Kanako Hirabayashi-Takahashi and Kuniaki Saito and Seiich Haga and Tomihiko Uemura and Susumu Izumi",
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Hirai, K, Kuroyanagi, H, Tatebayashi, Y, Hayashi, Y, Hirabayashi-Takahashi, K, Saito, K, Haga, S, Uemura, T & Izumi, S 2010, 'Dual role of the carboxyl-terminal region of pig liver l-kynurenine 3-monooxygenase: Mitochondrial-targeting signal and enzymatic activity', Journal of Biochemistry, vol. 148, no. 6, pp. 639-650. https://doi.org/10.1093/jb/mvq099

Dual role of the carboxyl-terminal region of pig liver l-kynurenine 3-monooxygenase : Mitochondrial-targeting signal and enzymatic activity. / Hirai, Kumiko; Kuroyanagi, Hidehito; Tatebayashi, Yoshitaka; Hayashi, Yoshitaka; Hirabayashi-Takahashi, Kanako; Saito, Kuniaki; Haga, Seiich; Uemura, Tomihiko; Izumi, Susumu.

In: Journal of Biochemistry, Vol. 148, No. 6, 01.12.2010, p. 639-650.

Research output: Contribution to journalArticle

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T1 - Dual role of the carboxyl-terminal region of pig liver l-kynurenine 3-monooxygenase

T2 - Mitochondrial-targeting signal and enzymatic activity

AU - Hirai, Kumiko

AU - Kuroyanagi, Hidehito

AU - Tatebayashi, Yoshitaka

AU - Hayashi, Yoshitaka

AU - Hirabayashi-Takahashi, Kanako

AU - Saito, Kuniaki

AU - Haga, Seiich

AU - Uemura, Tomihiko

AU - Izumi, Susumu

PY - 2010/12/1

Y1 - 2010/12/1

N2 - l-kynurenine 3-monooxygenase (KMO) is an NAD(P)H-dependent flavin monooxygenase that catalyses the hydroxylation of l-kynurenine to 3-hydroxykynurenine, and is localized as an oligomer in the mitochondrial outer membrane. In the human brain, KMO may play an important role in the formation of two neurotoxins, 3-hydroxykynurenine and quinolinic acid, both of which provoke severe neurodegenerative diseases. In mosquitos, it plays a role in the formation both of eye pigment and of an exflagellation-inducing factor (xanthurenic acid). Here, we present evidence that the C-terminal region of pig liver KMO plays a dual role. First, it is required for the enzymatic activity. Second, it functions as a mitochondrial targeting signal as seen in monoamine oxidase B (MAO B) or outer membrane cytochrome b5. The first role was shown by the comparison of the enzymatic activity of two mutants (C-terminally FLAG-tagged KMO and carboxyl-terminal truncation form, KMOΔC50) with that of the wild-type enzyme expressed in COS-7 cells. The second role was demonstrated with fluorescence microscopy by the comparison of the intracellular localization of the wild-type, three carboxyl-terminal truncated forms (ΔC20, ΔC30 and ΔC50), C-terminally FLAG-tagged wild-type and a mutant KMO, where two arginine residues, Arg461-Arg462, were replaced with Ser residues.

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