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. Author manuscript; available in PMC: 2010 Apr 1.
Published in final edited form as: Biochim Biophys Acta. 2009 Jan 21;1791(4):314–320. doi: 10.1016/j.bbalip.2009.01.004

Fig. 1.

Fig. 1

Tafazzin exchanges acyl groups between PC and CL. A, Rat liver mitochondria (1.5 mg protein) were incubated for 30 min in 0.2 mL buffer with 0.1 μCi 16:0-[14C]18:2-PC and, where indicated, CL (10 nmol), bromoenol lactone (BEL, 20 μM), and methyl arachidonoyl fluorophosphonate (MAFP, 10 μM). Phospholipids were separated by two-dimensional thin-layer chromatography and stained with iodine. [14C]CL was isolated to measure radioactivity by liquid scintillation counting. B, Experiment was performed as above, except that purified Drosophila tafazzin (MBP-TAZ, 2.5 μg protein) was added instead of mitochondria. LPC (5 nmol) was added to one set of samples. Tracer PC and CL (10 nmol each) were added before chromatography for better visualization. C, Transacylation rates of purified MBP-TAZ (2.5 μg) were measured by 2 min incubations, either in the presence of 10 μM [14C]16:0-LPC and variable concentrations of 16:0-18:2-PE (circles) or in the presence of 10 μM 16:0-[14C]18:2-PE and variable concentrations of 16:0-LPC (triangles). D, Transacylation rates of purified MBP-TAZ (2.5 μg) were measured by 2 min incubations either with 10 μM [14C]16:0-LPC and 50 μM CL or with 10 μM 16:0-[14C]18:2-PC and 50 μM MLCL. E, Mechanism by which tafazzin (enzyme, E) exchanges acyl groups between CL and PC in the absence of free lysophospholipids. Red color indicates radioactive species in reference to the experiment of panel B. Bar graphs represent means ± SEM (n=3).