FIG 5.

LipL-dependent lipoyl relay between H proteins. Lipoyl transfer from LA-GcvH (10 μM) to GcvH-L (20 μM) (lanes 5 and 6) and from LA-GcvH-L (40 μM) to GcvH (20 μM) (lanes 7 and 8), with LipL (1 μM) (lanes 6 and 8) and without LipL (lanes 5 and 7), was assessed by immunoblotting with rabbit anti-lipoic acid antibody. Apo- (lanes 1 and 3) and LplA1-lipoylated (lanes 2 and 4) GcvH and GcvH-L (20 μM) were included as controls. A portion (2.5 μl) of a 50-μl reaction mixture was loaded into each lane. The presented blot is representative of at least three independent experiments.