Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers

Abstract Chimeric proteins have been widely used to evaluate intracellular protein–protein interactions (PPIs) in living cells with various readouts. By combining an interleukin-3-dependent murine cells and chimeric proteins containing a receptor tyrosine kinase c-kit, we previously established a c-...

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Autores principales:	Daiki Kashima, Masahiro Kawahara
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/31208666015c4a82ac498f5f0e6794b9
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spelling	oai:doaj.org-article:31208666015c4a82ac498f5f0e6794b92021-12-02T18:24:55ZEvolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers10.1038/s41598-021-91287-z2045-2322https://doaj.org/article/31208666015c4a82ac498f5f0e6794b92021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91287-zhttps://doaj.org/toc/2045-2322Abstract Chimeric proteins have been widely used to evaluate intracellular protein–protein interactions (PPIs) in living cells with various readouts. By combining an interleukin-3-dependent murine cells and chimeric proteins containing a receptor tyrosine kinase c-kit, we previously established a c-kit-based PPI screening (KIPPIS) system to evaluate and select protein binders. In the KIPPIS components, proteins of interest are connected with a chemically inducible helper module and the intracellular domain of the growth-signaling receptor c-kit, which detects PPIs based on cell proliferation as a readout. In this system, proteins of interest can be incorporated into chimeric proteins without any scaffold proteins, which would be advantageous for evaluating interaction between small peptides/domains. To prove this superiority, we apply KIPPIS to 6 peptide aptamer–polypeptide pairs, which are derived from endogenous, synthetic, and viral proteins. Consequently, all of the 6 peptide aptamer–polypeptide interactions are successfully detected by cell proliferation. The detection sensitivity can be modulated in a helper ligand-dependent manner. The assay results of KIPPIS correlate with the activation levels of Src, which is located downstream of c-kit-mediated signal transduction. Control experiments reveal that KIPPIS clearly discriminates interacting aptamers from non-interacting ones. Thus, KIPPIS proves to be a versatile platform for evaluating the binding properties of peptide aptamers.Daiki KashimaMasahiro KawaharaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Daiki Kashima Masahiro Kawahara Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers
description	Abstract Chimeric proteins have been widely used to evaluate intracellular protein–protein interactions (PPIs) in living cells with various readouts. By combining an interleukin-3-dependent murine cells and chimeric proteins containing a receptor tyrosine kinase c-kit, we previously established a c-kit-based PPI screening (KIPPIS) system to evaluate and select protein binders. In the KIPPIS components, proteins of interest are connected with a chemically inducible helper module and the intracellular domain of the growth-signaling receptor c-kit, which detects PPIs based on cell proliferation as a readout. In this system, proteins of interest can be incorporated into chimeric proteins without any scaffold proteins, which would be advantageous for evaluating interaction between small peptides/domains. To prove this superiority, we apply KIPPIS to 6 peptide aptamer–polypeptide pairs, which are derived from endogenous, synthetic, and viral proteins. Consequently, all of the 6 peptide aptamer–polypeptide interactions are successfully detected by cell proliferation. The detection sensitivity can be modulated in a helper ligand-dependent manner. The assay results of KIPPIS correlate with the activation levels of Src, which is located downstream of c-kit-mediated signal transduction. Control experiments reveal that KIPPIS clearly discriminates interacting aptamers from non-interacting ones. Thus, KIPPIS proves to be a versatile platform for evaluating the binding properties of peptide aptamers.
format	article
author	Daiki Kashima Masahiro Kawahara
author_facet	Daiki Kashima Masahiro Kawahara
author_sort	Daiki Kashima
title	Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers
title_short	Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers
title_full	Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers
title_fullStr	Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers
title_full_unstemmed	Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers
title_sort	evolution of kippis as a versatile platform for evaluating intracellularly functional peptide aptamers
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/31208666015c4a82ac498f5f0e6794b9
work_keys_str_mv	AT daikikashima evolutionofkippisasaversatileplatformforevaluatingintracellularlyfunctionalpeptideaptamers AT masahirokawahara evolutionofkippisasaversatileplatformforevaluatingintracellularlyfunctionalpeptideaptamers
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Evolution of KIPPIS as a versatile platform for evaluating intracellularly functional peptide aptamers

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