FIG. 9.
Cerulenin enhances the need for ATP and C26:0 in the microsomal assay. X2180 wt cells were precultured for 180 min with cerulenin (10 μg/ml) and for the last 90 min with myriocin (40 μg/ml) in addition (bars 3 to 9) or only with myriocin for 90 min (bars 1 and 2). Microsomal remodeling assays were run in duplicate under standard conditions (bars 1 and 3) or with ingredients omitted or added as indicated at the bottom. The amount of radioactivity in anchor peptides was determined by scintillation counting and plotted as a percentage of incorporation under standard conditions (bar 1). Reaction 6 contained 0.5 U of apyrase in addition, and 2 or 10 nanomoles of C26:0-CoA was added in conjunction with 0.1 mg of purified yeast acyl-CoA binding protein.