Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases.

Fiche publication

Date publication

avril 2025

Journal

STAR protocols

Auteurs

Membres identifiés du Cancéropôle Est :
Pr WEISSMAN Kira

Tous les auteurs :
Gusenda C, Berlage S, Burkhart I, Chagot B, Weissman KJ, Schwalbe H, Grininger M

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/40220306

Résumé

Acyl carrier proteins (ACPs) are key domains in fatty acid and polyketide synthases (PKSs), shuttling intermediates to catalytic partners. Here, we present a protocol for purifying and analyzing ACPs from fatty acid synthases (FASs) and PKSs, using four model ACPs from Mus musculus, Saccharopolyspora erythraea, Streptomyces venezuelae, and Streptomyces hygroscopicus. We describe steps for recombinant ACP production in E. coli; purification via chromatography or precipitation; ACP modification (holo/acyl forms); and analysis using urea PAGE, high-performance liquid chromatography (HPLC), and NMR spectroscopy. For complete details on the use and execution of this protocol, please refer to Gusenda et al..