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. 2011 Jul 29;286(39):33819–33831. doi: 10.1074/jbc.M111.269779

FIGURE 9.

FIGURE 9.

Determination of the site of acylation of 1-acyllyso-PG. Solubilized membranes prepared from induced BLR(DE3)pLysS/pET15b or BLR(DE3)pLysS/At1g78690p were incubated in an in vitro assay using 1-oleoyllyso-PG as the acyl acceptor and arachidonyl-CoA as the acyl donor as described under “Experimental Procedures.” The lipids from the reaction mixture were extracted and analyzed using ESI-MS. A, negative ion ESI-MS from m/z 700 to 800 of lipids extracted from the reaction of solubilized membranes from BLR(DE3)pLysS/pET15b. B, negative ion ESI-MS from m/z 700 to 800 of lipids extracted from the reaction of solubilized membranes from BLR(DE3)pLysS/pAt1g78690p. The mass spectra in A and B are of the material eluting between minutes 16.5 and 19.5 of the normal phase chromatography. C, negative ion MS/MS of the in vitro product at m/z 795.5. D, positive ion MS/MS of the in vitro product at m/z 797.5. E, positive ion MS/MS of 16:0, 18:1 PG standard. F, positive ion MS/MS of 3,1′-BMP. The insets show the major product ions predicted from each predicted precursor ion.