Figure 6.

LPAAT activities in E. coli membrane preparations. E. coli JC201 cells were transformed in pBluescript SK vector (Stratagene) with the BAT2 sequence and in vector only. Membranes were obtained from cells induced by IPTG addition to cultures grown at 30°C. The membranes were assayed in the presence of 50 mm Tris-HCl, pH 8, 1 mm MgCl₂, 55 μm LPA, 1 mm DTT, and varying concentrations of [1-¹⁴C]palmitoyl-CoA or [1-¹⁴C]oleoyl-oA, and the products of the reaction were resolved on TLC. The phosphatidic acid spot was visualized by autoradiography and collected for scintillation counting. Values are the means obtained with two individual membrane preparations. The values obtained with E. coli JC201/pBSK for palmitoyl-CoA and oleoyl-CoA were similar and are labeled as acyl-CoA.