Gräwe, Alexander ; Ranglack, Jan ; Weyrich, Anastasia ; Stein, Viktor (2021)
Synthetic protein switches: Combinatorial linker engineering with iFLinkC.
In: Methods in enzymology, 647
doi: 10.1016/bs.mie.2020.09.009
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
Linker engineering constitutes a critical, yet frequently underestimated aspect in the construction of synthetic protein switches and sensors. Notably, systematic strategies to engineer linkers by predictive means remain largely elusive to date. This is primarily due to our insufficient understanding how the biophysical properties that underlie linker functions mediate the conformational transitions in artificially engineered protein switches and sensors. The construction of synthetic protein switches and sensors therefore heavily relies on experimental trial-and-error. Yet, methods for effectively generating linker diversity at the genetic level are scarce. Addressing this technical shortcoming, iterative functional linker cloning (iFLinkC) enables the combinatorial assembly of linker elements with functional domains from sequence verified repositories that are developed and stored in-house. The assembly process is highly scalable and given its recursive nature generates linker diversity in a combinatorial and exponential fashion based on a limited number of linker elements.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2021 |
| Autor(en): | Gräwe, Alexander ; Ranglack, Jan ; Weyrich, Anastasia ; Stein, Viktor |
| Art des Eintrags: | Bibliographie |
| Titel: | Synthetic protein switches: Combinatorial linker engineering with iFLinkC. |
| Sprache: | Englisch |
| Publikationsjahr: | Januar 2021 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Methods in enzymology |
| Jahrgang/Volume einer Zeitschrift: | 647 |
| DOI: | 10.1016/bs.mie.2020.09.009 |
| Kurzbeschreibung (Abstract): | Linker engineering constitutes a critical, yet frequently underestimated aspect in the construction of synthetic protein switches and sensors. Notably, systematic strategies to engineer linkers by predictive means remain largely elusive to date. This is primarily due to our insufficient understanding how the biophysical properties that underlie linker functions mediate the conformational transitions in artificially engineered protein switches and sensors. The construction of synthetic protein switches and sensors therefore heavily relies on experimental trial-and-error. Yet, methods for effectively generating linker diversity at the genetic level are scarce. Addressing this technical shortcoming, iterative functional linker cloning (iFLinkC) enables the combinatorial assembly of linker elements with functional domains from sequence verified repositories that are developed and stored in-house. The assembly process is highly scalable and given its recursive nature generates linker diversity in a combinatorial and exponential fashion based on a limited number of linker elements. |
| ID-Nummer: | pmid:33482991 |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Protein Engineering of Ion Conducting Nanopores |
| Hinterlegungsdatum: | 25 Jan 2021 14:05 |
| Letzte Änderung: | 25 Jan 2021 14:05 |
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