Fig. 1.

Size-exclusion chromatography of proteolytic hydrolysates of HeLa cell total proteins on Sephadex LH-20. Cells were labeled for 36 hours with [5-³H]mevalonic acid in the presence of 30 µM mevinolin, harvested, washed with phosphate-buffered saline, and extracted with lipid solvents. Cell pellets were then successively digested with proteases, and labeled digestion products were concentrated and purified by step elution from DEAE Sephacel. The eluted material was then passed through Sephadex LH-20 in 20% formic acid in ethanol at a flow rate of 0.25 nl/mmin, and 1.0-ml fractions were collected (4). Peak A, which corresponded to 1000-dalton material, contained 74% of the recovered label. Peak B, which corresponded to 500-dalton material, contained 22% of the recovered label. Recovery of the total applied label was 78%. Comparable chromatograms were obtained in six separate experiments.