Mueller, Lena (2019)
Chemical synthesis of switchable peptide-based nanopores: from ion channels to bio-inspired materials.
Technische Universität Darmstadt
doi: 10.25534/tuprints-00011495
Dissertation, Erstveröffentlichung
Kurzbeschreibung (Abstract)
The generation of different types of peptide-based nanopores is elucidated in this work. First, biological nanopores are investigated. Ion channels and pore-forming proteins represent an ultra-selective and ultra-sensitive category of nanopores. Eukaryotic potassium channels are structures of enormous size while prokaryotic potassium channels are comparable in their overall gating behavior but some are extremely reduced in their size. Within this work, the total chemical synthesis of the viral potassium channel KcvNTS is shown using Solid Phase Peptide Synthesis (SPPS) and Native Chemical Ligation (NCL) towards product formation. The main attention is on overcoming the challenging key steps including the substantial insolubility of the extensively hydrophobic fragments. Secondly, hybrid nanopores are generated. Nanopores composed of solid-state materials can be used as a robust and stable scaffold to integrate a selective and sensitive peptide moiety to generate a hybrid nanopore for a sensing application. In this work, the immobilization of the Amino Terminal Cu(II)- and Ni(II) binding motif (ATCUN) to conically shaped PET-based solid state nanopores is undertaken. The generated sensor is tuned (pH 6.5) to only bind Cu(II) leading to the design of a selective and sensitive Cu(II)-sensor (limit of detection in solution 13.5 nM using fluorescence titration; hybrid system in fM range using I-V measurements). Furthermore, the motif is investigated substantially regarding the mandatory histidine moiety. Mutants are designed and examined towards their binding behavior. Additionally, the DNA scission ability of the ATCUN motif and its mutants is investigated towards several plasmids paving the way for the design of a sequencing device using the presented polymer-based hybrid nanopore system.
| Typ des Eintrags: | Dissertation | ||||
|---|---|---|---|---|---|
| Erschienen: | 2019 | ||||
| Autor(en): | Mueller, Lena | ||||
| Art des Eintrags: | Erstveröffentlichung | ||||
| Titel: | Chemical synthesis of switchable peptide-based nanopores: from ion channels to bio-inspired materials | ||||
| Sprache: | Englisch | ||||
| Referenten: | Tietze, Asst. Prof Alesia A. ; Thiel, Prof. Dr. Gerhard | ||||
| Publikationsjahr: | 13 Dezember 2019 | ||||
| Ort: | Darmstadt | ||||
| Datum der mündlichen Prüfung: | 27 Februar 2020 | ||||
| DOI: | 10.25534/tuprints-00011495 | ||||
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/11495 | ||||
| Kurzbeschreibung (Abstract): | The generation of different types of peptide-based nanopores is elucidated in this work. First, biological nanopores are investigated. Ion channels and pore-forming proteins represent an ultra-selective and ultra-sensitive category of nanopores. Eukaryotic potassium channels are structures of enormous size while prokaryotic potassium channels are comparable in their overall gating behavior but some are extremely reduced in their size. Within this work, the total chemical synthesis of the viral potassium channel KcvNTS is shown using Solid Phase Peptide Synthesis (SPPS) and Native Chemical Ligation (NCL) towards product formation. The main attention is on overcoming the challenging key steps including the substantial insolubility of the extensively hydrophobic fragments. Secondly, hybrid nanopores are generated. Nanopores composed of solid-state materials can be used as a robust and stable scaffold to integrate a selective and sensitive peptide moiety to generate a hybrid nanopore for a sensing application. In this work, the immobilization of the Amino Terminal Cu(II)- and Ni(II) binding motif (ATCUN) to conically shaped PET-based solid state nanopores is undertaken. The generated sensor is tuned (pH 6.5) to only bind Cu(II) leading to the design of a selective and sensitive Cu(II)-sensor (limit of detection in solution 13.5 nM using fluorescence titration; hybrid system in fM range using I-V measurements). Furthermore, the motif is investigated substantially regarding the mandatory histidine moiety. Mutants are designed and examined towards their binding behavior. Additionally, the DNA scission ability of the ATCUN motif and its mutants is investigated towards several plasmids paving the way for the design of a sequencing device using the presented polymer-based hybrid nanopore system. |
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| URN: | urn:nbn:de:tuda-tuprints-114951 | ||||
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 500 Naturwissenschaften 500 Naturwissenschaften und Mathematik > 540 Chemie |
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| Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie |
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| Hinterlegungsdatum: | 02 Jun 2020 07:31 | ||||
| Letzte Änderung: | 09 Jun 2020 06:18 | ||||
| PPN: | |||||
| Referenten: | Tietze, Asst. Prof Alesia A. ; Thiel, Prof. Dr. Gerhard | ||||
| Datum der mündlichen Prüfung / Verteidigung / mdl. Prüfung: | 27 Februar 2020 | ||||
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| Suche nach Titel in: | TUfind oder in Google | ||||
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