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Figure 3 | BMC Biochemistry

Figure 3

From: Acyl-CoA-binding and self-associating properties of a recombinant 13.3 kDa N-terminal fragment of diacylglycerol acyltransferase-1 from oilseed rape

Figure 3

Effect of CoA on binding of acyl-CoA to BnDGAT1(1–116)His6 and demonstration of acyl-CoA binding using electrophoresis. Panel A: Effect of non-radiolabeled CoA (8 μM) or oleoyl-CoA (8 μM) on the binding of [1-14C]oleoyl-CoA to BnDGAT1(1–116)His6. Each incubation mixture contained 0.2 μM protein and 19 μM radiolabeled acyl-CoA. Following sedimentation of Lipidex-1000, equal aliquots of the supernatants were assayed for radioactivity. Each incubation was performed in triplicate and each data point represents the mean ± SD. Panel B: Binding of acyl-CoA to BnDGAT1(1–116)His6 demonstrated using non-denaturing gel electrophoresis and phosphorimaging. Phosphorimage obtained using radiolabeled erucoyl-CoA (lane 1) and BnDGAT1(1–116)His6 incubated with radiolabeled erucoyl-CoA (lane 2). Protein stain of BnDGAT1(1–116)His6 incubated with radiolabeled erucoyl-CoA is shown in lane 3.

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