Bacterial fermentation and isotope labelling optimized for amyloidogenic proteins.

ABSTRACT: We developed a cost sensitive isotope labelling procedure using a fed-batch fermentation method and tested its efficiency producing the ¹⁵ N-, ¹³ C- and ¹⁵ N/¹³ C-labelled variants of an amyloidogenic miniprotein (E5: EEEAVRLYIQWLKEGGPSSGRPPPS). E5 is a surface active protein, which forms amyloids in solution. Here, we confirm, using both PM-IRRAS and AFM measurements, that the air-water interface triggers structural rearrangement and promotes the amyloid formation of E5, and thus it is a suitable test protein to work out efficient isotope labelling schemes even for such difficult sequences. E. coli cells expressing the recombinant, ubiquitin-fused miniprotein were grown in minimal media containing either unlabelled nutrients, or ¹⁵ N-

SUBMITTER: Vida I 

PROVIDER: S-EPMC8085922 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature