Vypal2: a versatile peptide ligase for precision tailoring of proteins

The last two decades have seen an increasing demand for new protein-modification methods from the biotech industry and biomedical research communities. Owing to their mild aqueous reaction conditions, enzymatic methods based on the use of peptide ligases are particularly desirable. In this regard, t...

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Main Authors: Zhang, Dingpeng, Wang, Zhen, Hu, Side, Lescar, Julien, Tam, James P., Liu, Chuan-Fa
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2022
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Online Access:https://hdl.handle.net/10356/160205
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-1602052023-02-28T17:10:32Z Vypal2: a versatile peptide ligase for precision tailoring of proteins Zhang, Dingpeng Wang, Zhen Hu, Side Lescar, Julien Tam, James P. Liu, Chuan-Fa School of Biological Sciences Science::Biological sciences Biocatalysts Protein Labeling The last two decades have seen an increasing demand for new protein-modification methods from the biotech industry and biomedical research communities. Owing to their mild aqueous reaction conditions, enzymatic methods based on the use of peptide ligases are particularly desirable. In this regard, the recently discovered peptidyl Asx-specific ligases (PALs) have emerged as powerful biotechnological tools in recent years. However, as a new class of peptide ligases, their scope and application remain underexplored. Herein, we report the use of a new PAL, VyPAL2, for a diverse range of protein modifications. We successfully showed that VyPAL2 was an efficient biocatalyst for protein labelling, inter-protein ligation, and protein cyclization. The labelled or cyclized protein ligands remained functionally active in binding to their target receptors. We also demonstrated on-cell labelling of protein ligands pre-bound to cellular receptors and cell-surface engineering via modifying a covalently anchored peptide substrate pre-installed on cell-surface glycans. Together, these examples firmly establish Asx-specific ligases, such as VyPAL2, as the biocatalysts of the future for site-specific protein modification, with a myriad of applications in basic research and drug discovery. Ministry of Education (MOE) Published version This research was supported by Academic Research Grant Tier 3 (MOE2016-T3-1-003) from the Singapore Ministry of Education (MOE) to the J.P.T., J.L. and C.-F.L. laboratories, and by AcRF Tier 1 (2019-T1-002-100) from MOE to C.-F.L. 2022-07-15T05:49:26Z 2022-07-15T05:49:26Z 2022 Journal Article Zhang, D., Wang, Z., Hu, S., Lescar, J., Tam, J. P. & Liu, C. (2022). Vypal2: a versatile peptide ligase for precision tailoring of proteins. International Journal of Molecular Sciences, 23(1), 458-. https://dx.doi.org/10.3390/ijms23010458 1661-6596 https://hdl.handle.net/10356/160205 10.3390/ijms23010458 35008882 2-s2.0-85122045585 1 23 458 en MOE2016-T3-1-003 MOE2019-T1-002-100 International Journal of Molecular Sciences © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
Biocatalysts
Protein Labeling
spellingShingle Science::Biological sciences
Biocatalysts
Protein Labeling
Zhang, Dingpeng
Wang, Zhen
Hu, Side
Lescar, Julien
Tam, James P.
Liu, Chuan-Fa
Vypal2: a versatile peptide ligase for precision tailoring of proteins
description The last two decades have seen an increasing demand for new protein-modification methods from the biotech industry and biomedical research communities. Owing to their mild aqueous reaction conditions, enzymatic methods based on the use of peptide ligases are particularly desirable. In this regard, the recently discovered peptidyl Asx-specific ligases (PALs) have emerged as powerful biotechnological tools in recent years. However, as a new class of peptide ligases, their scope and application remain underexplored. Herein, we report the use of a new PAL, VyPAL2, for a diverse range of protein modifications. We successfully showed that VyPAL2 was an efficient biocatalyst for protein labelling, inter-protein ligation, and protein cyclization. The labelled or cyclized protein ligands remained functionally active in binding to their target receptors. We also demonstrated on-cell labelling of protein ligands pre-bound to cellular receptors and cell-surface engineering via modifying a covalently anchored peptide substrate pre-installed on cell-surface glycans. Together, these examples firmly establish Asx-specific ligases, such as VyPAL2, as the biocatalysts of the future for site-specific protein modification, with a myriad of applications in basic research and drug discovery.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Zhang, Dingpeng
Wang, Zhen
Hu, Side
Lescar, Julien
Tam, James P.
Liu, Chuan-Fa
format Article
author Zhang, Dingpeng
Wang, Zhen
Hu, Side
Lescar, Julien
Tam, James P.
Liu, Chuan-Fa
author_sort Zhang, Dingpeng
title Vypal2: a versatile peptide ligase for precision tailoring of proteins
title_short Vypal2: a versatile peptide ligase for precision tailoring of proteins
title_full Vypal2: a versatile peptide ligase for precision tailoring of proteins
title_fullStr Vypal2: a versatile peptide ligase for precision tailoring of proteins
title_full_unstemmed Vypal2: a versatile peptide ligase for precision tailoring of proteins
title_sort vypal2: a versatile peptide ligase for precision tailoring of proteins
publishDate 2022
url https://hdl.handle.net/10356/160205
_version_ 1759857927943880704