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Development of a cellular tau enzyme-linked immunosorbent assay method for screening GSK-3β inhibitors.
Assay Drug Dev Technol. 2011 Oct; 9(5):503-13.AD

Abstract

Glycogen synthase kinase-3β (GSK-3β), a serine/threonine kinase also known as tau protein kinase I, has been implicated in the pathogenic conditions of Alzheimer's disease. Many investigators have focused on GSK-3 inhibitor as a therapeutic drug. In this study, we established a cell-based assay for the screening of novel GSK-3β inhibitors. For this purpose, four-repeat tau cDNAs were stably expressed in human embryonic kidney 293 (HEK293) cells (HEK293-Tau). The proliferation of HEK293-Tau cells was no different from that of HEK293 cells, as measured by the bromodeoxyuridine enzyme-linked immunosorbent assay (BrdU ELISA). The concentration-dependent reduction of tau phosphorylation by GSK-3 inhibitors, LiCl, Chir98023, and SB415286, was examined by immunoblot analysis and Tau ELISA (in situ ELISA). Highly consistent data were obtained, suggesting that this novel ELISA method is highly reproducible. Using this ELISA strategy, we isolated a few candidate compounds, including compounds 114 and 149, from several hundreds of synthetic agents and demonstrated that such candidates protect nerve growth factor-differentiated PC12 cells against amyloid-β-induced cell death. These data indicate that this Tau ELISA method in HEK293-Tau cells may be a suitable cell-based assay system to screen for GSK-3β inhibitors.

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Authors+Show Affiliations

Cho GW
Department of Neurology, College of Medicine, Hanyang University, #17 Haengdang-dong, Seongdong-gu, Seoul, Korea.
Noh MY
No affiliation info available
Kang BY
No affiliation info available
Ku IW
No affiliation info available
Park J
No affiliation info available
Hong YH
No affiliation info available
Kim MH
No affiliation info available
Kim SH
No affiliation info available

MeSH

Alzheimer DiseaseAminophenolsAnimalsCell Culture TechniquesCell SurvivalDose-Response Relationship, DrugDrug DiscoveryDrug Evaluation, PreclinicalEnzyme InhibitorsEnzyme-Linked Immunosorbent AssayGlycogen Synthase Kinase 3HEK293 CellsHumansImidazolesImmunoblottingMaleimidesMolecular Targeted TherapyNeuroprotective AgentsPC12 CellsPhosphorylationPlasmidsPyridinesPyrimidinesRatstau Proteins

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21561378

Citation

Cho, Goang-Won, et al. "Development of a Cellular Tau Enzyme-linked Immunosorbent Assay Method for Screening GSK-3β Inhibitors." Assay and Drug Development Technologies, vol. 9, no. 5, 2011, pp. 503-13.
Cho GW, Noh MY, Kang BY, et al. Development of a cellular tau enzyme-linked immunosorbent assay method for screening GSK-3β inhibitors. Assay Drug Dev Technol. 2011;9(5):503-13.
Cho, G. W., Noh, M. Y., Kang, B. Y., Ku, I. W., Park, J., Hong, Y. H., Kim, M. H., & Kim, S. H. (2011). Development of a cellular tau enzyme-linked immunosorbent assay method for screening GSK-3β inhibitors. Assay and Drug Development Technologies, 9(5), 503-13. https://doi.org/10.1089/adt.2010.0343
Cho GW, et al. Development of a Cellular Tau Enzyme-linked Immunosorbent Assay Method for Screening GSK-3β Inhibitors. Assay Drug Dev Technol. 2011;9(5):503-13. PubMed PMID: 21561378.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a cellular tau enzyme-linked immunosorbent assay method for screening GSK-3β inhibitors. AU - Cho,Goang-Won, AU - Noh,Min-Young, AU - Kang,Byung Yong, AU - Ku,Il-Whea, AU - Park,Jiseon, AU - Hong,Yoon-Ho, AU - Kim,Myung-Hwa, AU - Kim,Seung Hyun, Y1 - 2011/05/11/ PY - 2011/5/13/entrez PY - 2011/5/13/pubmed PY - 2012/4/20/medline SP - 503 EP - 13 JF - Assay and drug development technologies JO - Assay Drug Dev Technol VL - 9 IS - 5 N2 - Glycogen synthase kinase-3β (GSK-3β), a serine/threonine kinase also known as tau protein kinase I, has been implicated in the pathogenic conditions of Alzheimer's disease. Many investigators have focused on GSK-3 inhibitor as a therapeutic drug. In this study, we established a cell-based assay for the screening of novel GSK-3β inhibitors. For this purpose, four-repeat tau cDNAs were stably expressed in human embryonic kidney 293 (HEK293) cells (HEK293-Tau). The proliferation of HEK293-Tau cells was no different from that of HEK293 cells, as measured by the bromodeoxyuridine enzyme-linked immunosorbent assay (BrdU ELISA). The concentration-dependent reduction of tau phosphorylation by GSK-3 inhibitors, LiCl, Chir98023, and SB415286, was examined by immunoblot analysis and Tau ELISA (in situ ELISA). Highly consistent data were obtained, suggesting that this novel ELISA method is highly reproducible. Using this ELISA strategy, we isolated a few candidate compounds, including compounds 114 and 149, from several hundreds of synthetic agents and demonstrated that such candidates protect nerve growth factor-differentiated PC12 cells against amyloid-β-induced cell death. These data indicate that this Tau ELISA method in HEK293-Tau cells may be a suitable cell-based assay system to screen for GSK-3β inhibitors. SN - 1557-8127 UR - https://www.unboundmedicine.com/medline/citation/21561378/Development_of_a_cellular_tau_enzyme_linked_immunosorbent_assay_method_for_screening_GSK_3β_inhibitors_ L2 - https://www.liebertpub.com/doi/10.1089/adt.2010.0343?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -