A high throughput approach for the generation of orthogonally interacting protein pairs

Abstract In contrast to the nearly error-free self-assembly of protein architectures in nature, artificial assembly of protein complexes with pre-defined structure and function in vitro is still challenging. To mimic nature’s strategy to construct pre-defined three-dimensional protein architectures,...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Justin Lawrie, Xi Song, Wei Niu, Jiantao Guo
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a6e8eaecf7b8478c82664f701c2535e5
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a6e8eaecf7b8478c82664f701c2535e5
record_format	dspace
spelling	oai:doaj.org-article:a6e8eaecf7b8478c82664f701c2535e52021-12-02T15:08:51ZA high throughput approach for the generation of orthogonally interacting protein pairs10.1038/s41598-018-19281-62045-2322https://doaj.org/article/a6e8eaecf7b8478c82664f701c2535e52018-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-19281-6https://doaj.org/toc/2045-2322Abstract In contrast to the nearly error-free self-assembly of protein architectures in nature, artificial assembly of protein complexes with pre-defined structure and function in vitro is still challenging. To mimic nature’s strategy to construct pre-defined three-dimensional protein architectures, highly specific protein-protein interacting pairs are needed. Here we report an effort to create an orthogonally interacting protein pair from its parental pair using a bacteria-based in vivo directed evolution strategy. This high throughput approach features a combination of a negative and a positive selection. The newly developed negative selection from this work was used to remove any protein mutants that retain effective interaction with their parents. The positive selection was used to identify mutant pairs that can engage in effective mutual interaction. By using the cohesin-dockerin protein pair that is responsible for the self-assembly of cellulosome as a model system, we demonstrated that a protein pair that is orthogonal to its parent pair could be readily generated using our strategy. This approach could open new avenues to a wide range of protein-based assembly, such as biocatalysis or nanomaterials, with pre-determined architecture and potentially novel functions and properties.Justin LawrieXi SongWei NiuJiantao GuoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-9 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Justin Lawrie Xi Song Wei Niu Jiantao Guo A high throughput approach for the generation of orthogonally interacting protein pairs
description	Abstract In contrast to the nearly error-free self-assembly of protein architectures in nature, artificial assembly of protein complexes with pre-defined structure and function in vitro is still challenging. To mimic nature’s strategy to construct pre-defined three-dimensional protein architectures, highly specific protein-protein interacting pairs are needed. Here we report an effort to create an orthogonally interacting protein pair from its parental pair using a bacteria-based in vivo directed evolution strategy. This high throughput approach features a combination of a negative and a positive selection. The newly developed negative selection from this work was used to remove any protein mutants that retain effective interaction with their parents. The positive selection was used to identify mutant pairs that can engage in effective mutual interaction. By using the cohesin-dockerin protein pair that is responsible for the self-assembly of cellulosome as a model system, we demonstrated that a protein pair that is orthogonal to its parent pair could be readily generated using our strategy. This approach could open new avenues to a wide range of protein-based assembly, such as biocatalysis or nanomaterials, with pre-determined architecture and potentially novel functions and properties.
format	article
author	Justin Lawrie Xi Song Wei Niu Jiantao Guo
author_facet	Justin Lawrie Xi Song Wei Niu Jiantao Guo
author_sort	Justin Lawrie
title	A high throughput approach for the generation of orthogonally interacting protein pairs
title_short	A high throughput approach for the generation of orthogonally interacting protein pairs
title_full	A high throughput approach for the generation of orthogonally interacting protein pairs
title_fullStr	A high throughput approach for the generation of orthogonally interacting protein pairs
title_full_unstemmed	A high throughput approach for the generation of orthogonally interacting protein pairs
title_sort	high throughput approach for the generation of orthogonally interacting protein pairs
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/a6e8eaecf7b8478c82664f701c2535e5
work_keys_str_mv	AT justinlawrie ahighthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs AT xisong ahighthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs AT weiniu ahighthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs AT jiantaoguo ahighthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs AT justinlawrie highthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs AT xisong highthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs AT weiniu highthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs AT jiantaoguo highthroughputapproachforthegenerationoforthogonallyinteractingproteinpairs
_version_	1718388019095928832

A high throughput approach for the generation of orthogonally interacting protein pairs

Ejemplares similares