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. Author manuscript; available in PMC: 2014 Nov 19.
Published in final edited form as: Biochemistry. 2013 Nov 4;52(46):10.1021/bi401116n. doi: 10.1021/bi401116n

Figure 3. Reconstitution of acetyl-CoA carboxylase.

Figure 3

(A) SDS-PAGE analysis of the acetyl-CoA carboxylase subunits. AccA, holo-AccB, AccC and AccD. AccB migrates around 20 kDa in a Tris-glycine SDS-PAGE system, because of its long Pro- and Ala-rich segment between residues 34 and 1017. (B) Effect of acetyl-CoA concentration on the initial reaction velocity of the complete reconstituted system shown in Figure 1. Reaction condition: 30 μM AccABCD, 1 μM FabABDFGH, 10 μM FabIZ, 30 μM holo-ACP, 30 μM TesA, 15 mM bicarbonate, 1 mM ATP, 5 mM MgCl2, and 1.3 mM NADPH. (C) Dependence of overall turnover rate on acetyl-CoA carboxylase concentration in the complete reconstituted system. Reactions contain varying concentrations of the carboxylase complex, 1 μM FabABDFGH, 10 μM FabIZ, 30 μM holo-ACP, 30 μM TesA (TE), 600 μM acetyl-CoA, 15 mM sodium bicarbonate, 1 mM ATP, 5 mM MgCl2, and 1.3 mM NADPH.