Figure 8 | Scientific Reports

Figure 8

From: Phosphatidylserine decarboxylase CT699, lysophospholipid acyltransferase CT775, and acyl-ACP synthase CT776 provide membrane lipid diversity to Chlamydia trachomatis

Figure 8

Acyl-CoA::1-acyl-GPC acyltransferase activity of CT775. CT775 was produced in E. coli, and activity in the membrane fraction was measured as described previously30. Reactions were performed with 1 µM NBD-C16-CoA in the presence of 10 µM synthetic 1-C16-sn-glycerol-3-P-choline (>90% 1-acyl-GPC isomer) at 30 °C. NBD-PC obtained after 30 minutes reaction with 40 µg of CT775 microsomes was extracted and separated by TLC. The NBD-PC spot was scraped from the plate and treated with bee venom PLA2 (bvPLA2). Lipids were extracted and separated by TLC, and the fluorescence in LPC (1- or 2-acyl-GPC), FA and unreacted PC was quantified. Experiments were performed with CT775 (panel A) and human LPCAT1 (panel B). NBD-LPC was used as a migration standard; it is shown in the right-hand lane of the TLC plate in panel B.

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