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https://hdl.handle.net/10356/162124
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DC Field | Value | Language |
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dc.contributor.author | Zhang, Dingpeng | en_US |
dc.contributor.author | Wang, Zhen | en_US |
dc.contributor.author | Hu, Side | en_US |
dc.contributor.author | Chan, Ning-Yu | en_US |
dc.contributor.author | Liew, Heng Tai | en_US |
dc.contributor.author | Lescar, Julien | en_US |
dc.contributor.author | Tam, James P. | en_US |
dc.contributor.author | Liu, Chuan-Fa | en_US |
dc.date.accessioned | 2022-10-04T08:25:06Z | - |
dc.date.available | 2022-10-04T08:25:06Z | - |
dc.date.issued | 2022 | - |
dc.identifier.citation | Zhang, D., Wang, Z., Hu, S., Chan, N., Liew, H. T., Lescar, J., Tam, J. P. & Liu, C. (2022). Asparaginyl endopeptidase-mediated protein C-terminal hydrazinolysis for the synthesis of bioconjugates. Bioconjugate Chemistry, 33(1), 238-247. https://dx.doi.org/10.1021/acs.bioconjchem.1c00551 | en_US |
dc.identifier.issn | 1043-1802 | en_US |
dc.identifier.uri | https://hdl.handle.net/10356/162124 | - |
dc.description.abstract | Asparaginyl endopeptidases (AEPs) are cysteinyl enzymes naturally catalyzing the hydrolysis and transpeptidation reactions at Asx-Xaa bonds. These reactions go by a common acyl-enzyme thioester intermediate, which is either attacked by water (for a protease-AEP) or by a peptidic amine nucleophile (for a ligase-AEP) to form the respective hydrolysis or aminolysis product. Herein, we show that hydrazine and hydroxylamine, two α-effect nucleophiles, are capable of resolving the thioester intermediate to yield peptide and protein products containing a C-terminal hydrazide and hydroxamic acid functionality, respectively. The hydrazinolysis reaction exhibits very high efficiency and can be completed in minutes at a low enzyme-to-substrate ratio. We further show the utility of the so-formed asparaginyl hydrazide in native chemical ligation and hydrazone conjugation. Using an EGFR-targeting affibody as a model protein, we have showcased our methodology in the preparation of a number of protein ligation or conjugation products, which are decorated with various functional moieties. The ZEGFR affibody-doxorubicin conjugate shows high selective binding and cytotoxicity toward the EGFR-positive A431 cells. Our results demonstrate the advantages of AEP-mediated protein hydrazinolysis as a simple and straightforward strategy for the precision manufacturing of protein bioconjugates. | en_US |
dc.description.sponsorship | Ministry of Education (MOE) | en_US |
dc.language.iso | en | en_US |
dc.relation | MOE2016-T3-1-003 | en_US |
dc.relation | 2019-T1-002-100 | en_US |
dc.relation | NGF-2019-07-029 | en_US |
dc.relation.ispartof | Bioconjugate Chemistry | en_US |
dc.rights | © 2022 American Chemical Society. All rights reserved. | en_US |
dc.subject | Science::Biological sciences | en_US |
dc.title | Asparaginyl endopeptidase-mediated protein C-terminal hydrazinolysis for the synthesis of bioconjugates | en_US |
dc.type | Journal Article | en |
dc.contributor.school | School of Biological Sciences | en_US |
dc.identifier.doi | 10.1021/acs.bioconjchem.1c00551 | - |
dc.identifier.pmid | 34985285 | - |
dc.identifier.scopus | 2-s2.0-85122824481 | - |
dc.identifier.issue | 1 | en_US |
dc.identifier.volume | 33 | en_US |
dc.identifier.spage | 238 | en_US |
dc.identifier.epage | 247 | en_US |
dc.subject.keywords | Asparaginyl | en_US |
dc.subject.keywords | Bioconjugates | en_US |
dc.description.acknowledgement | 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) and NTUitive Gap grant (NGF-2019-07-029) from MOE to the C.-F.L. laboratory. | en_US |
item.fulltext | No Fulltext | - |
item.grantfulltext | none | - |
Appears in Collections: | SBS Journal Articles |
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