FIGURE 10.

Tandem affinity-purified YLR290C-CNAP co-purifies with Q and late-stage Q intermediates. For each strain, aliquots of the anti-PC eluates (29% of E1 and 29% of E2) were combined and subjected to lipid extraction with methanol and petroleum ether. Lipid extracts were analyzed by HPLC-MS/MS for total Q₆ (Q₆ + Q₆H₂) (A), DMQ₆ (C), and IDMQ₆ (E). Measured lipids were normalized by the extracted eluate volume. The bars represent the mean of two measurements, and the error bars represent the standard deviation. Statistical significance was determined with the two-tailed Student's t test, and the lowercase letters above the bars are indicative of statistical significance. In A, the content of Q₆ in CA-1 was 5.4-fold higher compared with wild type (a, p = 0.0418). In C, the content of DMQ₆ in CA-1 was 6.7-fold higher compared with wild type (a, p = 0.0025). In E, there was no detectable IDMQ₆ in the wild type. ND, not detected. Representative overlaid traces of both strains (W303, magenta; CA-1, green) are shown for Q₆ (B), DMQ₆ (D), and IDMQ₆ (F).