{"id":{"repo_id":"penn","oai_identifier":"oai:repository.upenn.edu:20.500.14332/28455"},"canonical_url":"https://search.dev.ndltd.org/etd/penn/oai:repository.upenn.edu:20.500.14332/28455","repository":{"repo_id":"penn","name":"University of Pennsylvania","base_url":"https://repository.upenn.edu/server/oai/request"},"display":{"title":"Stapling and Unstapling Peptides and Proteins With S-Tetrazine","abstract":"This thesis will focus on the design, synthesis, and validation of synthetic techniques to introduce s-tetrazine into peptides and proteins. The s-tetrazine molecule is effective for restricting peptides/proteins to macrocyclic conformations (i.e. stapling). Importantly, the incorporated s-tetrazine chromophore will undergo photodisassociation upon absorp-tion of a photon permitting the release of the restricted conformations (i.e. unstapling). In chapter one, we aimed to study a fundamental folding process known as the helix-coil transition by phototriggering coupled with transient two-dimensional infrared spec-troscopy (2D IR). The s-tetrazine molecule possesses the photochemical properties of an ideal phototrigger, as such, s-tetrazine was employed towards the development of tech-niques capable of capturing the fastest structural transitions of biomolecules with both high spatial and high temporal resolution. Tripeptide linchpins, containing the s-tetrazine phototrigger, were prepared by solid-phase peptide synthesis. The latter were then em-ployed toward the construction kinked helices near equilibrium via a fragment coupling procedure. The relaxation of the kinked helical structures were observed by pump/probe transient 2D IR spectroscopy. In chapter two, new synthetic protocols have been developed and validated for the in-troduction of s-tetrazine into peptides and proteins to staple and unstaple the confor-mations. Conditions for the introduction of s-tetrazine into cysteine sulfhydryl groups of unprotected peptides conducted with aqueous biphasic conditions, permitting the con-struction of macrocyclic peptides with a wide range of functionally and ring topology, bridging from one to 27 amino acid residues adjoining the cysteines. Importantly, the sta-pled conformations were released photochemically to their thiocyanate counterparts, and in turn the resulting thiocyanates removed to regenerate the native peptide. To the best of our knowledge s-tetrazine comprises the first example of a readily removable peptide sta-ple. Finally, the stapling and unstapling protocol has been extended to include thioredox-in as an example of a protein with an incorporated s-tetrazine construct that can also serve a useful role in conjugation strategies.","abstract_html":"This thesis will focus on the design, synthesis, and validation of synthetic techniques to introduce s-tetrazine into peptides and proteins. The s-tetrazine molecule is effective for restricting peptides/proteins to macrocyclic conformations (i.e. stapling). Importantly, the incorporated s-tetrazine chromophore will undergo photodisassociation upon absorp-tion of a photon permitting the release of the restricted conformations (i.e. unstapling). In chapter one, we aimed to study a fundamental folding process known as the helix-coil transition by phototriggering coupled with transient two-dimensional infrared spec-troscopy (2D IR). The s-tetrazine molecule possesses the photochemical properties of an ideal phototrigger, as such, s-tetrazine was employed towards the development of tech-niques capable of capturing the fastest structural transitions of biomolecules with both high spatial and high temporal resolution. Tripeptide linchpins, containing the s-tetrazine phototrigger, were prepared by solid-phase peptide synthesis. The latter were then em-ployed toward the construction kinked helices near equilibrium via a fragment coupling procedure. The relaxation of the kinked helical structures were observed by pump/probe transient 2D IR spectroscopy. In chapter two, new synthetic protocols have been developed and validated for the in-troduction of s-tetrazine into peptides and proteins to staple and unstaple the confor-mations. Conditions for the introduction of s-tetrazine into cysteine sulfhydryl groups of unprotected peptides conducted with aqueous biphasic conditions, permitting the con-struction of macrocyclic peptides with a wide range of functionally and ring topology, bridging from one to 27 amino acid residues adjoining the cysteines. Importantly, the sta-pled conformations were released photochemically to their thiocyanate counterparts, and in turn the resulting thiocyanates removed to regenerate the native peptide. To the best of our knowledge s-tetrazine comprises the first example of a readily removable peptide sta-ple. Finally, the stapling and unstapling protocol has been extended to include thioredox-in as an example of a protein with an incorporated s-tetrazine construct that can also serve a useful role in conjugation strategies.","abstract_has_math":false,"creators":["Brown, Stephen P"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Amos B. Smith"],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015-01-01","date_published":"2015-01-01","updated_at":"2026-07-24T03:45:15Z","subjects":[],"languages":["en"],"rights":["Stephen P. Brown"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://repository.upenn.edu/handle/20.500.14332/28455","outbound_label":"Repository record","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Amos B. 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The s-tetrazine molecule possesses the photochemical properties of an ideal phototrigger, as such, s-tetrazine was employed towards the development of tech-niques capable of capturing the fastest structural transitions of biomolecules with both high spatial and high temporal resolution. Tripeptide linchpins, containing the s-tetrazine phototrigger, were prepared by solid-phase peptide synthesis. The latter were then em-ployed toward the construction kinked helices near equilibrium via a fragment coupling procedure. The relaxation of the kinked helical structures were observed by pump/probe transient 2D IR spectroscopy. In chapter two, new synthetic protocols have been developed and validated for the in-troduction of s-tetrazine into peptides and proteins to staple and unstaple the confor-mations. Conditions for the introduction of s-tetrazine into cysteine sulfhydryl groups of unprotected peptides conducted with aqueous biphasic conditions, permitting the con-struction of macrocyclic peptides with a wide range of functionally and ring topology, bridging from one to 27 amino acid residues adjoining the cysteines. Importantly, the sta-pled conformations were released photochemically to their thiocyanate counterparts, and in turn the resulting thiocyanates removed to regenerate the native peptide. To the best of our knowledge s-tetrazine comprises the first example of a readily removable peptide sta-ple. Finally, the stapling and unstapling protocol has been extended to include thioredox-in as an example of a protein with an incorporated s-tetrazine construct that can also serve a useful role in conjugation strategies."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Doctor of Philosophy (PhD)"]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Stapling and Unstapling Peptides and Proteins With S-Tetrazine"]}]}],"canonical_facts":{"dc:contributor.advisor":["Amos B. Smith"],"dc:creator":["Brown, Stephen P"],"dc:date":["2023-05-17T15:52:14.000"],"dc:date.accessioned":["2023-05-22T16:41:56Z"],"dc:date.available":["2018-10-11T00:00:00Z"],"dc:date.issued":["2015-01-01"],"dc:description.abstract":["This thesis will focus on the design, synthesis, and validation of synthetic techniques to introduce s-tetrazine into peptides and proteins. The s-tetrazine molecule is effective for restricting peptides/proteins to macrocyclic conformations (i.e. stapling). Importantly, the incorporated s-tetrazine chromophore will undergo photodisassociation upon absorp-tion of a photon permitting the release of the restricted conformations (i.e. unstapling). In chapter one, we aimed to study a fundamental folding process known as the helix-coil transition by phototriggering coupled with transient two-dimensional infrared spec-troscopy (2D IR). The s-tetrazine molecule possesses the photochemical properties of an ideal phototrigger, as such, s-tetrazine was employed towards the development of tech-niques capable of capturing the fastest structural transitions of biomolecules with both high spatial and high temporal resolution. Tripeptide linchpins, containing the s-tetrazine phototrigger, were prepared by solid-phase peptide synthesis. The latter were then em-ployed toward the construction kinked helices near equilibrium via a fragment coupling procedure. The relaxation of the kinked helical structures were observed by pump/probe transient 2D IR spectroscopy. In chapter two, new synthetic protocols have been developed and validated for the in-troduction of s-tetrazine into peptides and proteins to staple and unstaple the confor-mations. Conditions for the introduction of s-tetrazine into cysteine sulfhydryl groups of unprotected peptides conducted with aqueous biphasic conditions, permitting the con-struction of macrocyclic peptides with a wide range of functionally and ring topology, bridging from one to 27 amino acid residues adjoining the cysteines. Importantly, the sta-pled conformations were released photochemically to their thiocyanate counterparts, and in turn the resulting thiocyanates removed to regenerate the native peptide. To the best of our knowledge s-tetrazine comprises the first example of a readily removable peptide sta-ple. Finally, the stapling and unstapling protocol has been extended to include thioredox-in as an example of a protein with an incorporated s-tetrazine construct that can also serve a useful role in conjugation strategies."],"dc:description.degree":["Doctor of Philosophy (PhD)"],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["https://repository.upenn.edu/handle/20.500.14332/28455"],"dc:language":["en"],"dc:rights":["Stephen P. Brown"],"dc:title":["Stapling and Unstapling Peptides and Proteins With S-Tetrazine"],"dc:type":["Dissertation/Thesis"]},"updated_at":"2026-07-24T03:45:15Z"}