Supplementary Figure 2: Tethered catalysis baits used for PISA screening in yeast. | Nature Methods

Supplementary Figure 2: Tethered catalysis baits used for PISA screening in yeast.

From: Post-translational selective intracellular silencing of acetylated proteins with de novo selected intrabodies

Supplementary Figure 2

(a) Schematic representation of tethered catalysis baits for PISA screening. Target antigen is fused at the C-terminus of LexA DNA Binding Domain (DBD) and at the N-terminus of a Histone Acetyl Transferase (HAT) catalytic domain. As descried in Guo et al. (Nat. Biotechnol., 888–892 (2004)), the antigen is post-translationally modified in a constitutive manner by the wild type version of the enzyme (left, screening bait), but it is not modified when HAT is mutated with an inactivating single point mutation (right, control bait). (b) Histone H3 (H3) and Integrase (IN) baits were expressed in yeast and immunoprecipitated with anti-HA antibody (rat monoclonal). After IP, their expression was verified by Western Blot using anti-HA Ab (rabbit polyclonal). Acetylation has also been verified with either an anti-H3Ac antibody or an anti-acetyl-lysine antibody.

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