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Synthetic protein switches: Combinatorial linker engineering with iFLinkC.

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
Article, Bibliographie

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.

Item Type: Article
Erschienen: 2021
Creators: Gräwe, Alexander ; Ranglack, Jan ; Weyrich, Anastasia ; Stein, Viktor
Type of entry: Bibliographie
Title: Synthetic protein switches: Combinatorial linker engineering with iFLinkC.
Language: English
Date: January 2021
Journal or Publication Title: Methods in enzymology
Volume of the journal: 647
DOI: 10.1016/bs.mie.2020.09.009
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.

Identification Number: pmid:33482991
Divisions: 10 Department of Biology
10 Department of Biology > Protein Engineering of Ion Conducting Nanopores
Date Deposited: 25 Jan 2021 14:05
Last Modified: 25 Jan 2021 14:05
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