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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Autores: Sánchez Campillo, Iván, Blanco Canosa, Juan B.
Tipo de recurso: artículo
Fecha de publicación:2024
País:España
Institución: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
Acceso en línea:https://hdl.handle.net/2445/224424
Access Level:acceso abierto
Palabra clave: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 Peptides Sánchez Campillo, Iván Blanco Canosa, Juan B. Cinètica química Síntesi proteica Pèptids Chemical kinetics Protein synthesis 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 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 Publications https://hdl.handle.net/2445/224424
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
author Sánchez Campillo, Iván
author_facet Sánchez Campillo, Iván
Blanco Canosa, Juan B.
author_role author
author2 Blanco Canosa, Juan B.
author2_role author
topic Cinètica química
Síntesi proteica
Pèptids
Chemical kinetics
Protein synthesis
Peptides
topic_facet 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
format article
url https://hdl.handle.net/2445/224424
eu_rights_str_mv openAccess
publisher ACS Publications
institution Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
collection Recercat. Dipósit de la Recerca de Catalunya
reponame_str Recercat. Dipósit de la Recerca de Catalunya
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
_version_ 1878441721101025280
publishDateSort 2024
author_browse Blanco Canosa, Juan B.
Sánchez Campillo, Iván
publisherStr ACS Publications
score 6.924472

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