Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system

We report expression and purification of a FLT3 protein with ITD mutation (FLT3-ITD) with a steady tyrosine kinase activity using a silkworm-baculovirus system, and its application as a fast screening system of tyrosine kinase inhibitors. The FLT3-ITD protein was expressed in Bombyx mori L. pupae in...

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Autores principales: Yamamoto, Naoki, Kikuchi, Jiro, Furukawa, Yusuke, Shibayama, Naoya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9070948/
https://www.ncbi.nlm.nih.gov/pubmed/35511790
http://dx.doi.org/10.1371/journal.pone.0261699
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author Yamamoto, Naoki
Kikuchi, Jiro
Furukawa, Yusuke
Shibayama, Naoya
author_facet Yamamoto, Naoki
Kikuchi, Jiro
Furukawa, Yusuke
Shibayama, Naoya
author_sort Yamamoto, Naoki
collection PubMed
description We report expression and purification of a FLT3 protein with ITD mutation (FLT3-ITD) with a steady tyrosine kinase activity using a silkworm-baculovirus system, and its application as a fast screening system of tyrosine kinase inhibitors. The FLT3-ITD protein was expressed in Bombyx mori L. pupae infected by gene-modified nucleopolyhedrovirus, and was purified as an active state. We performed an inhibition assay using 17 kinase inhibitors, and succeeded in screening two inhibitors for FLT3-ITD. The result has paved the way for screening FLT3-ITD inhibitors in a fast and easy manner, and also for structural studies.
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spelling pubmed-90709482022-05-06 Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system Yamamoto, Naoki Kikuchi, Jiro Furukawa, Yusuke Shibayama, Naoya PLoS One Research Article We report expression and purification of a FLT3 protein with ITD mutation (FLT3-ITD) with a steady tyrosine kinase activity using a silkworm-baculovirus system, and its application as a fast screening system of tyrosine kinase inhibitors. The FLT3-ITD protein was expressed in Bombyx mori L. pupae infected by gene-modified nucleopolyhedrovirus, and was purified as an active state. We performed an inhibition assay using 17 kinase inhibitors, and succeeded in screening two inhibitors for FLT3-ITD. The result has paved the way for screening FLT3-ITD inhibitors in a fast and easy manner, and also for structural studies. Public Library of Science 2022-05-05 /pmc/articles/PMC9070948/ /pubmed/35511790 http://dx.doi.org/10.1371/journal.pone.0261699 Text en © 2022 Yamamoto et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Yamamoto, Naoki
Kikuchi, Jiro
Furukawa, Yusuke
Shibayama, Naoya
Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system
title Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system
title_full Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system
title_fullStr Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system
title_full_unstemmed Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system
title_short Fast in-vitro screening of FLT3-ITD inhibitors using silkworm-baculovirus protein expression system
title_sort fast in-vitro screening of flt3-itd inhibitors using silkworm-baculovirus protein expression system
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9070948/
https://www.ncbi.nlm.nih.gov/pubmed/35511790
http://dx.doi.org/10.1371/journal.pone.0261699
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