Supplementary Figure 1: Purification of the native ribosome–NatA complex.
From: Ribosome–NatA architecture reveals that rRNA expansion segments coordinate N-terminal acetylation
a, Schematic representation of the construct for TAP-tagged Naa15. CaMBD, calmodulin-binding domain; TEV, tobacco etch virus protease cleavage site; ProtA, protein A IgG binding domains. b, Amido black–stained PVDF membrane after transfer of purification samples separated on a 4–12% NuPAGE gel. c, Western blot of purification samples probing with an antibody directed against the CaMBD (anti-CAB, Thermo Fisher) and against uL29 (polyclonal chicken antiserum, Davids Biotechnolgie). L, lysate; FT, flow through; R, resuspension; W, wash; E, elution; B, boiled beads. Naa15-CaMBD corresponds to TEV-cleaved product carrying a calmodulin-binding domain, which is recognized by the anti-CAB antibody. 0.1 A260 of E was loaded, for L and FT 1/40,000 and for wash fractions 1/7 of the volume was loaded on the gel.
