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  SNaPe: a versatile method to generate multiplexed protein fusions using synthetic linker peptides for in vitro applications

Ulrich, V., & Cryle, M. (2017). SNaPe: a versatile method to generate multiplexed protein fusions using synthetic linker peptides for in vitro applications. Journal of Peptide Science, 23(1), 16-27. doi:10.1002/psc.2943.

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 Creators:
Ulrich, Veronika1, Author           
Cryle, Max1, Author           
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1Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Max Planck Society, ou_1497700              

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Free keywords: peptide; protein; ligation; fusion; biosynthetic enzymes
 Abstract: Understanding the structure and function of protein complexes and multi-domain proteins is highly important in biology, although the in vitro characterization of these systems is often complicated by their size or the transient nature of protein/protein interactions. To assist in the characterization of such protein complexes, we have developed a modular approach to fusion protein generation that relies upon Sortase-mediated and Native chemical ligation using synthetic Peptide linkers (SNaPe) to link two separately expressed proteins. In this approach, we utilize two separate linking steps – sortase-mediated and native chemical ligation – together with a library of peptide linkers to generate libraries of fusion proteins. We have demonstrated the viability of SNaPe to generate libraries from fusion protein constructs taken from the biosynthetic enzymes responsible for late stage aglycone assembly during glycopeptide antibiotic biosynthesis. Crucially, SNaPe was able to generate fusion proteins that are inaccessible via direct expression of the fusion construct itself. This highlights the advantages of SNaPe to not only access fusion proteins that have been previously unavailable for biochemical and structural characterization but also to do so in a manner that enables the linker itself to be controlled as an experimental parameter of fusion protein generation. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

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Language(s): eng - English
 Dates: 2016-10-262016-09-092016-11-062016-12-022017-01-01
 Publication Status: Issued
 Pages: 12
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/psc.2943
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Title: Journal of Peptide Science
  Other : J. Peptide Sci.
Source Genre: Journal
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Publ. Info: Chichester, West Sussex, UK : John Wiley & Sons
Pages: - Volume / Issue: 23 (1) Sequence Number: - Start / End Page: 16 - 27 Identifier: ISSN: 1075-2617
CoNE: https://pure.mpg.de/cone/journals/resource/954925604784