HIF2 ODD-luciferase reporter: the most sensitive assay for HIF prolyl hydroxylase inhibitors
- Авторлар: Smirnova N.1, Osipyants A.1, Khristichenko A.1, Hushpulian D.1, Nikulin S.1, Chubar T.2, Zakhariants A.3, Tishkov V.2,3, Gazaryan I.1,2, Poloznikov A.1
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Мекемелер:
- D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology
- Department of Chemical Enzymology, Faculty of Chemistry, M. V. Lomonosov Moscow State University
- Innovation and High Technologies MSU Ltd
- Шығарылым: Том 67, № 1 (2018)
- Беттер: 150-156
- Бөлім: Article
- URL: https://journals.rcsi.science/1066-5285/article/view/242007
- DOI: https://doi.org/10.1007/s11172-018-2051-5
- ID: 242007
Дәйексөз келтіру
Аннотация
Reporters expressing fusion proteins of HIF2 and HIF3 C-terminal oxygen degradable domain (ODD) with the firefly luciferase, HIF2 ODD-luc and HIF3 ODD-luc, were constructed and briefly characterized. Stable neuroblastoma cell lines expressing either reporter were generated, and their response to the known HIF prolyl hydroxylase inhibitors: dimethyloxalylglycine, ciclopirox, and adaptaquin, was studied and compared with the HIF1 ODD-luc reporter. The HIF2 ODD-luc reporter exhibited the highest sensitivity: its response in absolute luminescence value was almost an order of magnitude higher than that of the HIF1 ODD-luc reporter. The new reporter can be used for a fine discrimination of enzyme inhibitors stabilizing HIF2, and further structural optimization of adaptaquin discovered earlier by using the HIF1 ODD-luc reporter. The higher sensitivity of HIF2 ODD-luc reporter could be most likely explained by the lower affinity of the endogenous enzyme for this HIF isoform in comparison with the two others, which also resulted in the increased efficiency of inhibitors under the reporter assay conditions.
Негізгі сөздер
Авторлар туралы
N. Smirnova
D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 1 ul. Samory Mashela, Moscow, 117997
A. Osipyants
D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 1 ul. Samory Mashela, Moscow, 117997
A. Khristichenko
D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 1 ul. Samory Mashela, Moscow, 117997
D. Hushpulian
D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 1 ul. Samory Mashela, Moscow, 117997
S. Nikulin
D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 1 ul. Samory Mashela, Moscow, 117997
T. Chubar
Department of Chemical Enzymology, Faculty of Chemistry, M. V. Lomonosov Moscow State University
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, Build. 3, 1 Leninskie Gory, Moscow, 119991
A. Zakhariants
Innovation and High Technologies MSU Ltd
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 16 ul. Tsimlyanskaya, Moscow, 109451
V. Tishkov
Department of Chemical Enzymology, Faculty of Chemistry, M. V. Lomonosov Moscow State University; Innovation and High Technologies MSU Ltd
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, Build. 3, 1 Leninskie Gory, Moscow, 119991; 16 ul. Tsimlyanskaya, Moscow, 109451
I. Gazaryan
D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology; Department of Chemical Enzymology, Faculty of Chemistry, M. V. Lomonosov Moscow State University
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 1 ul. Samory Mashela, Moscow, 117997; Build. 3, 1 Leninskie Gory, Moscow, 119991
A. Poloznikov
D. Rogachev Federal Scientific and Clinical Center of Pediatric Hematology, Oncology, and Immunology
Хат алмасуға жауапты Автор.
Email: andrey.poloznikov@fccho-moscow.ru
Ресей, 1 ul. Samory Mashela, Moscow, 117997