Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase

This study reports the application of expanding genetic codes in developing protein cage-based delivery systems. The evolved Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS)•tRNAPyl pairs derived from directed evolution are examined to probe their recognition for para-substituted phenylalanin...

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Autores principales:	Yi-Hui Wang, Mu-Lung Jian, Pei-Jung Chen, Jo-Chu Tsou, Le P. Truong, Yane-Shih Wang
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	pyrrolysyl-tRNA synthetase ferritin-drug conjugates strain-promoted azide-alkyne cycloaddition p-Azido-l-phenylalanine amber suppression efficiency drug delivery Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/cd39a17a669f493f9cd3cfbd2b0f9a5a
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spelling	oai:doaj.org-article:cd39a17a669f493f9cd3cfbd2b0f9a5a2021-12-01T00:09:43ZFerritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase2296-264610.3389/fchem.2021.779976https://doaj.org/article/cd39a17a669f493f9cd3cfbd2b0f9a5a2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fchem.2021.779976/fullhttps://doaj.org/toc/2296-2646This study reports the application of expanding genetic codes in developing protein cage-based delivery systems. The evolved Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS)•tRNAPyl pairs derived from directed evolution are examined to probe their recognition for para-substituted phenylalanine analogs. The evolved MmPylRS, AzFRS, harboring a wide range of substrates, is further engineered at the C-terminal region into another variant, AzFRS-MS. AzFRS-MS shows suppression of the elevated sfGFP protein amount up to 10 TAG stop codons when charging p-azido-l-phenylalanine (AzF, 4), which allows the occurrence of click chemistry. Since protein nanocages used as drug delivery systems that encompass multiple drugs through a site-specific loading approach remain largely unexplored, as a proof of concept, the application of AzFRS-MS for the site-specific incorporation of AzF on human heavy chain ferritin (Ftn) is developed. The Ftn-4 conjugate is shown to be able to load multiple fluorescence dyes or a therapeutic agent, doxorubicin (Dox), through the strain-promoted azide-alkyne cycloaddition (SPAAC) click reaction. Aiming to selectively target Her2+ breast cancer cells, Ftn-4-DOX conjugates fused with a HER2 receptor recognition peptide, anti-Her2/neu peptide (AHNP), is developed and demonstrated to be able to deliver Dox into the cell and to prolong the drug release. This work presents another application of evolved MmPylRS systems, whose potential in developing a variety of protein conjugates is noteworthy.Yi-Hui WangMu-Lung JianMu-Lung JianPei-Jung ChenPei-Jung ChenJo-Chu TsouJo-Chu TsouLe P. TruongYane-Shih WangYane-Shih WangFrontiers Media S.A.articlepyrrolysyl-tRNA synthetaseferritin-drug conjugatesstrain-promoted azide-alkyne cycloadditionp-Azido-l-phenylalanineamber suppression efficiencydrug deliveryChemistryQD1-999ENFrontiers in Chemistry, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	pyrrolysyl-tRNA synthetase ferritin-drug conjugates strain-promoted azide-alkyne cycloaddition p-Azido-l-phenylalanine amber suppression efficiency drug delivery Chemistry QD1-999
spellingShingle	pyrrolysyl-tRNA synthetase ferritin-drug conjugates strain-promoted azide-alkyne cycloaddition p-Azido-l-phenylalanine amber suppression efficiency drug delivery Chemistry QD1-999 Yi-Hui Wang Mu-Lung Jian Mu-Lung Jian Pei-Jung Chen Pei-Jung Chen Jo-Chu Tsou Jo-Chu Tsou Le P. Truong Yane-Shih Wang Yane-Shih Wang Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase
description	This study reports the application of expanding genetic codes in developing protein cage-based delivery systems. The evolved Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS)•tRNAPyl pairs derived from directed evolution are examined to probe their recognition for para-substituted phenylalanine analogs. The evolved MmPylRS, AzFRS, harboring a wide range of substrates, is further engineered at the C-terminal region into another variant, AzFRS-MS. AzFRS-MS shows suppression of the elevated sfGFP protein amount up to 10 TAG stop codons when charging p-azido-l-phenylalanine (AzF, 4), which allows the occurrence of click chemistry. Since protein nanocages used as drug delivery systems that encompass multiple drugs through a site-specific loading approach remain largely unexplored, as a proof of concept, the application of AzFRS-MS for the site-specific incorporation of AzF on human heavy chain ferritin (Ftn) is developed. The Ftn-4 conjugate is shown to be able to load multiple fluorescence dyes or a therapeutic agent, doxorubicin (Dox), through the strain-promoted azide-alkyne cycloaddition (SPAAC) click reaction. Aiming to selectively target Her2+ breast cancer cells, Ftn-4-DOX conjugates fused with a HER2 receptor recognition peptide, anti-Her2/neu peptide (AHNP), is developed and demonstrated to be able to deliver Dox into the cell and to prolong the drug release. This work presents another application of evolved MmPylRS systems, whose potential in developing a variety of protein conjugates is noteworthy.
format	article
author	Yi-Hui Wang Mu-Lung Jian Mu-Lung Jian Pei-Jung Chen Pei-Jung Chen Jo-Chu Tsou Jo-Chu Tsou Le P. Truong Yane-Shih Wang Yane-Shih Wang
author_facet	Yi-Hui Wang Mu-Lung Jian Mu-Lung Jian Pei-Jung Chen Pei-Jung Chen Jo-Chu Tsou Jo-Chu Tsou Le P. Truong Yane-Shih Wang Yane-Shih Wang
author_sort	Yi-Hui Wang
title	Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase
title_short	Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase
title_full	Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase
title_fullStr	Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase
title_full_unstemmed	Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase
title_sort	ferritin conjugates with multiple clickable amino acids encoded by c-terminal engineered pyrrolysyl-trna synthetase
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/cd39a17a669f493f9cd3cfbd2b0f9a5a
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Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase

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