Site-Specific Labelling of Multidomain Proteins by Amber Codon Suppression.

ABSTRACT: The access to information on the dynamic behaviour of large proteins is usually hindered as spectroscopic methods require the site-specific attachment of biophysical probes. A powerful emerging tool to tackle this issue is amber codon suppression. Till date, its application on large and complex multidomain proteins of MDa size has not been reported. Herein, we systematically investigate the feasibility to introduce different non-canonical amino acids into a 540 kDa homodimeric fatty acid synthase type I by genetic code expansion with subsequent fluorescent labelling. Our approach relies on a microplate-based reporter assay of low complexity using a GFP fusion protein to quickly screen for sufficient suppression conditions. Once identified, these findings were successfully utilized to upsca

SUBMITTER: Heil CS 

PROVIDER: S-EPMC6173736 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature