A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors

Abstract Fibroblast growth factor receptors (FGFRs) generate various transduction signals by interaction with fibroblast growth factors (FGFs) and are involved in various biological functions such as cell proliferation, migration, and differentiation. Malfunction of these proteins may lead to the de...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Dae-Eun Cheong, Hye-Ji Choi, Su-Kyoung Yoo, Hun-Dong Lee, Geun-Joong Kim
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1052c16b7f064dbda9e425a7265849d3
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:1052c16b7f064dbda9e425a7265849d3
record_format	dspace
spelling	oai:doaj.org-article:1052c16b7f064dbda9e425a7265849d32021-11-08T10:48:54ZA designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors10.1038/s41598-021-01029-42045-2322https://doaj.org/article/1052c16b7f064dbda9e425a7265849d32021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01029-4https://doaj.org/toc/2045-2322Abstract Fibroblast growth factor receptors (FGFRs) generate various transduction signals by interaction with fibroblast growth factors (FGFs) and are involved in various biological functions such as cell proliferation, migration, and differentiation. Malfunction of these proteins may lead to the development of various diseases, including cancer. Accordingly, FGFRs are considered an alternative therapeutic target for protein and/or gene therapy. However, the screening of antagonists or agonists of FGFRs is challenging due to their complex structural features associated with protein expression. Herein, we conducted the development of a protease-free cleavable tag (PFCT) for enhancing the solubility of difficult-to express protein by combining maltose-binding protein (MBP) and the C-terminal region of Npu intein. To validate the availability of the resulting tag for the functional production of extracellular domains of FGFRs (Ec_FGFRs), we performed fusion of PFCT with the N-terminus of Ec_FGFRs and analyzed the expression patterns. Almost all PFCT-Ec_FGFR fusion proteins were mainly detected in the soluble fraction except for Ec_FGFR4. Upon addition of the N-terminal region of Npu intein, approximately 85% of the PFCT-Ec_FGFRs was separated into PFCT and Ec_FGFR via intein-mediated cleavage. Additionally, the structural integrity of Ec_FGFR was confirmed by affinity purification using heparin column. Taken together, our study demonstrated that the PFCT could be used for soluble expression and selective separation of Ec_FGFRs.Dae-Eun CheongHye-Ji ChoiSu-Kyoung YooHun-Dong LeeGeun-Joong KimNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Dae-Eun Cheong Hye-Ji Choi Su-Kyoung Yoo Hun-Dong Lee Geun-Joong Kim A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors
description	Abstract Fibroblast growth factor receptors (FGFRs) generate various transduction signals by interaction with fibroblast growth factors (FGFs) and are involved in various biological functions such as cell proliferation, migration, and differentiation. Malfunction of these proteins may lead to the development of various diseases, including cancer. Accordingly, FGFRs are considered an alternative therapeutic target for protein and/or gene therapy. However, the screening of antagonists or agonists of FGFRs is challenging due to their complex structural features associated with protein expression. Herein, we conducted the development of a protease-free cleavable tag (PFCT) for enhancing the solubility of difficult-to express protein by combining maltose-binding protein (MBP) and the C-terminal region of Npu intein. To validate the availability of the resulting tag for the functional production of extracellular domains of FGFRs (Ec_FGFRs), we performed fusion of PFCT with the N-terminus of Ec_FGFRs and analyzed the expression patterns. Almost all PFCT-Ec_FGFR fusion proteins were mainly detected in the soluble fraction except for Ec_FGFR4. Upon addition of the N-terminal region of Npu intein, approximately 85% of the PFCT-Ec_FGFRs was separated into PFCT and Ec_FGFR via intein-mediated cleavage. Additionally, the structural integrity of Ec_FGFR was confirmed by affinity purification using heparin column. Taken together, our study demonstrated that the PFCT could be used for soluble expression and selective separation of Ec_FGFRs.
format	article
author	Dae-Eun Cheong Hye-Ji Choi Su-Kyoung Yoo Hun-Dong Lee Geun-Joong Kim
author_facet	Dae-Eun Cheong Hye-Ji Choi Su-Kyoung Yoo Hun-Dong Lee Geun-Joong Kim
author_sort	Dae-Eun Cheong
title	A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors
title_short	A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors
title_full	A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors
title_fullStr	A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors
title_full_unstemmed	A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors
title_sort	designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1052c16b7f064dbda9e425a7265849d3
work_keys_str_mv	AT daeeuncheong adesignedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT hyejichoi adesignedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT sukyoungyoo adesignedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT hundonglee adesignedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT geunjoongkim adesignedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT daeeuncheong designedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT hyejichoi designedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT sukyoungyoo designedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT hundonglee designedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors AT geunjoongkim designedfusiontagforsolubleexpressionandselectiveseparationofextracellulardomainsoffibroblastgrowthfactorreceptors
_version_	1718442588115042304

A designed fusion tag for soluble expression and selective separation of extracellular domains of fibroblast growth factor receptors

Ejemplares similares