Co-immunoprecipitation of AtNAA50 binding partners

In humans and plants, 40 % of the proteome is co-translationally acetylated at the N-terminus by a single Nα-acetyltransferase (Nat) termed NatA. The core NatA complex consists of the catalytic subunit NAA10 and the ribosome-anchoring subunit NAA15. The regulatory subunit HYPK and the acetyltransferase NAA50 join this complex in humans. Even though both are conserved in Arabidopsis thaliana, only AtHYPK is known to interact with AtNatA. In order to verify the interaction between AtNAA50 and the core NatA subunits and to identify novel AtNAA50 interactors, wildtype leaf protein extracts were mixed with a specific antiserum against AtNAA50 (Armbruster et al., 2020). Subsequently, protein A/G beads were used to precipitate antibody-bound AtNAA50 and its interaction partners. As a negative control, protein A/G beads were incubated with leaf protein extracts lacking the specific antibody against AtNAA50.