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An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation
Amino‐terminal acetylation is a ubiquitous protein modification affecting the majority of eukaryote proteins to regulate stability and function. We describe an optimised recombinant expression system for rapid production of amino terminal‐acetylated proteins within bacteria. We go on to describe the...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5396276/ https://www.ncbi.nlm.nih.gov/pubmed/28214355 http://dx.doi.org/10.1002/1873-3468.12597 |
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author | Eastwood, Tara A. Baker, Karen Brooker, Holly R. Frank, Stefanie Mulvihill, Daniel P. |
author_facet | Eastwood, Tara A. Baker, Karen Brooker, Holly R. Frank, Stefanie Mulvihill, Daniel P. |
author_sort | Eastwood, Tara A. |
collection | PubMed |
description | Amino‐terminal acetylation is a ubiquitous protein modification affecting the majority of eukaryote proteins to regulate stability and function. We describe an optimised recombinant expression system for rapid production of amino terminal‐acetylated proteins within bacteria. We go on to describe the system's use in a fluorescence based in vivo assay for use in the high‐throughput screen to identify drugs that impact amino‐terminal acetylation‐dependent oligomerisation. These new tools and protocols will allow researchers to enhance routine recombinant protein production and identify new molecules for use in research and clinical applications. |
format | Online Article Text |
id | pubmed-5396276 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-53962762017-04-25 An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation Eastwood, Tara A. Baker, Karen Brooker, Holly R. Frank, Stefanie Mulvihill, Daniel P. FEBS Lett Research Letters Amino‐terminal acetylation is a ubiquitous protein modification affecting the majority of eukaryote proteins to regulate stability and function. We describe an optimised recombinant expression system for rapid production of amino terminal‐acetylated proteins within bacteria. We go on to describe the system's use in a fluorescence based in vivo assay for use in the high‐throughput screen to identify drugs that impact amino‐terminal acetylation‐dependent oligomerisation. These new tools and protocols will allow researchers to enhance routine recombinant protein production and identify new molecules for use in research and clinical applications. John Wiley and Sons Inc. 2017-03-06 2017-03 /pmc/articles/PMC5396276/ /pubmed/28214355 http://dx.doi.org/10.1002/1873-3468.12597 Text en © 2017 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Letters Eastwood, Tara A. Baker, Karen Brooker, Holly R. Frank, Stefanie Mulvihill, Daniel P. An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation |
title | An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation |
title_full | An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation |
title_fullStr | An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation |
title_full_unstemmed | An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation |
title_short | An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation |
title_sort | enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation |
topic | Research Letters |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5396276/ https://www.ncbi.nlm.nih.gov/pubmed/28214355 http://dx.doi.org/10.1002/1873-3468.12597 |
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