Figure 8

N-terminal acetyltransferase activity of the hNaa16p-hNaa10p complex. HeLa cells were transfected with phNAA16-FLAG and phNAA10-V5, and immunoprecipitated with anti-FLAG (+) or a negative control antibody (-). The immunoprecipitated hNaa16p-hNaa10p complex was subjected to an in vitro N-acetylation assay by the addition of acetylation buffer, a synthetic oligopeptide, [¹⁴-C]-Acetyl coenzyme A. The radioactivity incorporated into the peptide was determined by scintillation counting. The peptides used are [H]SESSSKSRWGRPVGRRRRPVRVYP [OH] (SESS) and [H]SPTPPLFRWGRPVGRRRRPVRVYP [OH] (SPTP). An equal level of immunoprecipitated material (input enzyme) was verified by Western-blotting analysis. Results are presented as mean ± SD from three independent experiments.