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An enhanced recombinant amino‐terminal acetylation system and novel in vivo high‐throughput screen for molecules affecting α‐synuclein oligomerisation
Author(s) -
Eastwood Tara A.,
Baker Karen,
Brooker Holly R.,
Frank Stefanie,
Mulvihill Daniel P.
Publication year - 2017
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.12597
Subject(s) - acetylation , recombinant dna , eukaryote , in vivo , biochemistry , chemistry , amino acid , high throughput screening , computational biology , microbiology and biotechnology , biology , gene , genome , genetics
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.