Construction of novel repeat proteins with rigid and predictable structures using a shared helix method

Abstract Generating artificial protein assemblies with complex shapes requires a method for connecting protein components with stable and predictable structures. Currently available methods for creating rigid protein assemblies rely on either complicated calculations or extensive trial and error. We...

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Autores principales:	Suk-Jun Youn, Na-Young Kwon, Ji Hyun Lee, Jin Hong Kim, Jinwoo Choi, Hayyoung Lee, Jie-Oh Lee
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ec86a6a197e44045a69a1f748c81db3a
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spelling	oai:doaj.org-article:ec86a6a197e44045a69a1f748c81db3a2021-12-02T12:32:49ZConstruction of novel repeat proteins with rigid and predictable structures using a shared helix method10.1038/s41598-017-02803-z2045-2322https://doaj.org/article/ec86a6a197e44045a69a1f748c81db3a2017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02803-zhttps://doaj.org/toc/2045-2322Abstract Generating artificial protein assemblies with complex shapes requires a method for connecting protein components with stable and predictable structures. Currently available methods for creating rigid protein assemblies rely on either complicated calculations or extensive trial and error. We describe a simple and efficient method for connecting two proteins via a fused alpha helix that is formed by joining two preexisting helices into a single extended helix. Because the end-to-end ligation of helices does not guarantee the formation of a continuous helix, we superimposed 1–2 turns of pairs of connecting helices by using a molecular graphics program. Then, we chose amino acids from the two natural sequences that would stabilize the connecting helix. This “shared helix method” is highly efficient. All the designed proteins that could be produced in Escherichia coli were readily crystallized and had the expected fusion structures. To prove the usefulness of this method, we produced two novel repeat proteins by assembling several copies of natural or artificial proteins with alpha helices at both termini. Their crystal structures demonstrated the successful assembly of the repeating units with the intended curved shapes. We propose that this method could dramatically expand the available repertoire of natural repeat proteins.Suk-Jun YounNa-Young KwonJi Hyun LeeJin Hong KimJinwoo ChoiHayyoung LeeJie-Oh LeeNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Suk-Jun Youn Na-Young Kwon Ji Hyun Lee Jin Hong Kim Jinwoo Choi Hayyoung Lee Jie-Oh Lee Construction of novel repeat proteins with rigid and predictable structures using a shared helix method
description	Abstract Generating artificial protein assemblies with complex shapes requires a method for connecting protein components with stable and predictable structures. Currently available methods for creating rigid protein assemblies rely on either complicated calculations or extensive trial and error. We describe a simple and efficient method for connecting two proteins via a fused alpha helix that is formed by joining two preexisting helices into a single extended helix. Because the end-to-end ligation of helices does not guarantee the formation of a continuous helix, we superimposed 1–2 turns of pairs of connecting helices by using a molecular graphics program. Then, we chose amino acids from the two natural sequences that would stabilize the connecting helix. This “shared helix method” is highly efficient. All the designed proteins that could be produced in Escherichia coli were readily crystallized and had the expected fusion structures. To prove the usefulness of this method, we produced two novel repeat proteins by assembling several copies of natural or artificial proteins with alpha helices at both termini. Their crystal structures demonstrated the successful assembly of the repeating units with the intended curved shapes. We propose that this method could dramatically expand the available repertoire of natural repeat proteins.
format	article
author	Suk-Jun Youn Na-Young Kwon Ji Hyun Lee Jin Hong Kim Jinwoo Choi Hayyoung Lee Jie-Oh Lee
author_facet	Suk-Jun Youn Na-Young Kwon Ji Hyun Lee Jin Hong Kim Jinwoo Choi Hayyoung Lee Jie-Oh Lee
author_sort	Suk-Jun Youn
title	Construction of novel repeat proteins with rigid and predictable structures using a shared helix method
title_short	Construction of novel repeat proteins with rigid and predictable structures using a shared helix method
title_full	Construction of novel repeat proteins with rigid and predictable structures using a shared helix method
title_fullStr	Construction of novel repeat proteins with rigid and predictable structures using a shared helix method
title_full_unstemmed	Construction of novel repeat proteins with rigid and predictable structures using a shared helix method
title_sort	construction of novel repeat proteins with rigid and predictable structures using a shared helix method
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/ec86a6a197e44045a69a1f748c81db3a
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_version_	1718393953119633408

Construction of novel repeat proteins with rigid and predictable structures using a shared helix method

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