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Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation

Gliomas are common intracranial neoplasias in dogs. However, the underlying pathogenic mechanisms remain unclear. In humans, isocitrate dehydrogenase 2 (IDH2) is often mutated in gliomas. Although almost human IDH2 mutations have been identified at the Arg172 codon, few studies have reported structu...

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Autores principales: KAWAKAMI, Shota, OCHIAI, Kazuhiko, KATO, Yuiko, MICHISHITA, Masaki, HIRAMA, Hinako, OBARA, Ryo, AZAKAMI, Daigo, WATANABE, Masami, OMI, Toshinori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5797864/
https://www.ncbi.nlm.nih.gov/pubmed/29162772
http://dx.doi.org/10.1292/jvms.17-0362
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author KAWAKAMI, Shota
OCHIAI, Kazuhiko
KATO, Yuiko
MICHISHITA, Masaki
HIRAMA, Hinako
OBARA, Ryo
AZAKAMI, Daigo
WATANABE, Masami
OMI, Toshinori
author_facet KAWAKAMI, Shota
OCHIAI, Kazuhiko
KATO, Yuiko
MICHISHITA, Masaki
HIRAMA, Hinako
OBARA, Ryo
AZAKAMI, Daigo
WATANABE, Masami
OMI, Toshinori
author_sort KAWAKAMI, Shota
collection PubMed
description Gliomas are common intracranial neoplasias in dogs. However, the underlying pathogenic mechanisms remain unclear. In humans, isocitrate dehydrogenase 2 (IDH2) is often mutated in gliomas. Although almost human IDH2 mutations have been identified at the Arg172 codon, few studies have reported structural, functional or mutational information for canine IDH2. In this study, we cloned the full-length canine IDH2 (cIDH2) cDNA and substituted wild type Arg174 (cIDH2 WT: corresponding to R172 of human IDH2) with Lys (cIDH2 R174K). The cIDH2 WT and R174K proteins were overexpressed in HeLa cells, and their presence was confirmed using an anti-human IDH2-WT mAb (clone: KrMab-3) and an anti-IDH2-R172K mAb (clone: KMab-1). The IDH2 activity between cIDH2 WT and cIDH2 R174K transfectants was compared by measuring the production of NADH and NADPH. NADPH production was lower for cIDH2 R174K than that for cIDH2 WT transfectants. Finally, we detected increased expression of hypoxia inducible factor-1 alpha (HIF-1α) in cIDH2 R174K transfectants. This indicates that mutations at R174 can potentially induce carcinogenesis in canine somatic cells.
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spelling pubmed-57978642018-02-08 Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation KAWAKAMI, Shota OCHIAI, Kazuhiko KATO, Yuiko MICHISHITA, Masaki HIRAMA, Hinako OBARA, Ryo AZAKAMI, Daigo WATANABE, Masami OMI, Toshinori J Vet Med Sci Biochemistry Gliomas are common intracranial neoplasias in dogs. However, the underlying pathogenic mechanisms remain unclear. In humans, isocitrate dehydrogenase 2 (IDH2) is often mutated in gliomas. Although almost human IDH2 mutations have been identified at the Arg172 codon, few studies have reported structural, functional or mutational information for canine IDH2. In this study, we cloned the full-length canine IDH2 (cIDH2) cDNA and substituted wild type Arg174 (cIDH2 WT: corresponding to R172 of human IDH2) with Lys (cIDH2 R174K). The cIDH2 WT and R174K proteins were overexpressed in HeLa cells, and their presence was confirmed using an anti-human IDH2-WT mAb (clone: KrMab-3) and an anti-IDH2-R172K mAb (clone: KMab-1). The IDH2 activity between cIDH2 WT and cIDH2 R174K transfectants was compared by measuring the production of NADH and NADPH. NADPH production was lower for cIDH2 R174K than that for cIDH2 WT transfectants. Finally, we detected increased expression of hypoxia inducible factor-1 alpha (HIF-1α) in cIDH2 R174K transfectants. This indicates that mutations at R174 can potentially induce carcinogenesis in canine somatic cells. The Japanese Society of Veterinary Science 2017-11-21 2018-01 /pmc/articles/PMC5797864/ /pubmed/29162772 http://dx.doi.org/10.1292/jvms.17-0362 Text en ©2018 The Japanese Society of Veterinary Science This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Biochemistry
KAWAKAMI, Shota
OCHIAI, Kazuhiko
KATO, Yuiko
MICHISHITA, Masaki
HIRAMA, Hinako
OBARA, Ryo
AZAKAMI, Daigo
WATANABE, Masami
OMI, Toshinori
Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation
title Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation
title_full Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation
title_fullStr Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation
title_full_unstemmed Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation
title_short Functional alteration of canine isocitrate dehydrogenase 2 (IDH2) via an R174K mutation
title_sort functional alteration of canine isocitrate dehydrogenase 2 (idh2) via an r174k mutation
topic Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5797864/
https://www.ncbi.nlm.nih.gov/pubmed/29162772
http://dx.doi.org/10.1292/jvms.17-0362
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