Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides

Native chemical ligation (NCL) ligates two unprotected peptides in an aqueous buffer. One of the fragments features a C-terminal α-thioester functional group, and the second bears an N-terminal cysteine. The reaction mechanism depicts two steps: an intermolecular thiol–thioester exchange resulting i...

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Detalhes bibliográficos
Autores: Sánchez Campillo, Iván, Blanco Canosa, Juan B.
Formato: artículo
Fecha de publicación:2024
País:España
Recursos:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/224424
Acesso em linha:https://hdl.handle.net/2445/224424
Access Level:acceso abierto
Palavra-chave:Cinètica química
Síntesi proteica
Pèptids
Chemical kinetics
Protein synthesis
Peptides
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spelling Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester PeptidesSánchez Campillo, IvánBlanco Canosa, Juan B.Cinètica químicaSíntesi proteicaPèptidsChemical kineticsProtein synthesisPeptidesNative chemical ligation (NCL) ligates two unprotected peptides in an aqueous buffer. One of the fragments features a C-terminal α-thioester functional group, and the second bears an N-terminal cysteine. The reaction mechanism depicts two steps: an intermolecular thiol–thioester exchange resulting in a transient thioester, followed by an intramolecular S-to-N acyl shift to yield the final native peptide bond. Although this mechanism is well established, the direct observation of the transient thioester has been elusive because the fast intramolecular rearrangement prevents its accumulation. Here, the use of α-selenoester peptides allows a faster first reaction and an early buildup of the intermediate, enabling its quantification and the kinetic monitoring of the first and second steps. The results show a correlation between the steric hindrance in the α-thioester residue and the rearrangement rate. In bulky residues, the S-to-N acyl shift has a significant contribution to the overall reaction rate. This is particularly notable for valine and likely for other similar β-branched amino acids.ACS Publications202520252024info:eu-repo/semantics/articleinfo:eu-repo/semantics/publisherVersion9 p.application/pdfhttps://hdl.handle.net/2445/224424Articles publicats en revistes (Química Inorgànica i Orgànica)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésReproducció del document publicat a: https://doi.org/10.1021/jacsau.4c00705JACS Au 2024, vol. 4, pp. 4374−4382cc by-nc-nd (c) Sánchez Campillo, Iván et al., 2024http://creativecommons.org/licenses/by-nc-nd/3.0/es/info:eu-repo/semantics/openAccessoai:recercat.cat:2445/2244242026-05-29T05:05:01Z
dc.title.none.fl_str_mv Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
title Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
spellingShingle Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
Sánchez Campillo, Iván
Cinètica química
Síntesi proteica
Pèptids
Chemical kinetics
Protein synthesis
Peptides
title_short Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
title_full Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
title_fullStr Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
title_full_unstemmed Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
title_sort Kinetic and Mechanistic Studies of Native Chemical Ligation with Phenyl α-Selenoester Peptides
dc.creator.none.fl_str_mv Sánchez Campillo, Iván
Blanco Canosa, Juan B.
author Sánchez Campillo, Iván
author_facet Sánchez Campillo, Iván
Blanco Canosa, Juan B.
author_role author
author2 Blanco Canosa, Juan B.
author2_role author
dc.subject.none.fl_str_mv Cinètica química
Síntesi proteica
Pèptids
Chemical kinetics
Protein synthesis
Peptides
topic Cinètica química
Síntesi proteica
Pèptids
Chemical kinetics
Protein synthesis
Peptides
description Native chemical ligation (NCL) ligates two unprotected peptides in an aqueous buffer. One of the fragments features a C-terminal α-thioester functional group, and the second bears an N-terminal cysteine. The reaction mechanism depicts two steps: an intermolecular thiol–thioester exchange resulting in a transient thioester, followed by an intramolecular S-to-N acyl shift to yield the final native peptide bond. Although this mechanism is well established, the direct observation of the transient thioester has been elusive because the fast intramolecular rearrangement prevents its accumulation. Here, the use of α-selenoester peptides allows a faster first reaction and an early buildup of the intermediate, enabling its quantification and the kinetic monitoring of the first and second steps. The results show a correlation between the steric hindrance in the α-thioester residue and the rearrangement rate. In bulky residues, the S-to-N acyl shift has a significant contribution to the overall reaction rate. This is particularly notable for valine and likely for other similar β-branched amino acids.
publishDate 2024
dc.date.none.fl_str_mv 2024
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publisherVersion
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/224424
url https://hdl.handle.net/2445/224424
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1021/jacsau.4c00705
JACS Au 2024, vol. 4, pp. 4374−4382
dc.rights.none.fl_str_mv cc by-nc-nd (c) Sánchez Campillo, Iván et al., 2024
http://creativecommons.org/licenses/by-nc-nd/3.0/es/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc by-nc-nd (c) Sánchez Campillo, Iván et al., 2024
http://creativecommons.org/licenses/by-nc-nd/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 9 p.
application/pdf
dc.publisher.none.fl_str_mv ACS Publications
publisher.none.fl_str_mv ACS Publications
dc.source.none.fl_str_mv Articles publicats en revistes (Química Inorgànica i Orgànica)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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