Fig. 2

Ccw12-β-lactamase fusion proteins. (A) The predicted 3D structure of fusion reporter protein Ccw12-β-B. The 3D structure is colour-coded according to the plDDT score and labelled to highlight structural features. The tracks on the right correlate these features with their locations in the primary sequence and their corresponding plDDT scores. (B) Map of a yeast shuttle vector encoding Ccw12-β-lactamase fusion proteins. Amp^R – gene conferring ampicillin resistance to E. coli, ori – E. coli plasmid replication origin, 2µ – yeast 2µ plasmid replication origin, HIS3 – yeast selective marker allowing his3Δ strains growth on media lacking histidine, PHO5 promoter – yeast promoter induced in phosphate-free medium, Ccw12-bla – open reading frame encoding Ccw12-β-lactamase fusion protein, CYC1 terminator – terminator of yeast CYC1 gene. (C) Anti-HA immunoblotting of Ccw12- and Pir2-fusion proteins. The immunoblot shows non-covalently and covalently bound Ccw12- and Pir2-fusion proteins. The lower membrane, stained after electrotransfer with Ponceau S, serves as a loading control. (D) Relative β-lactamase activities of Ccw12- and Pir2-fusion proteins. The graph displays the activities normalised to Ccw12-β-A, with error bars indicating standard deviations. *p-value < 0.05, **p-value < 10^− 6