Figure 5 | Scientific Reports

Figure 5

From: The Multiple DSF-family QS Signals are Synthesized from Carbohydrate and Branched-chain Amino Acids via the FAS Elongation Cycle

Figure 5

The effect of RpfF on acyl-ACP thioesters.

(a) SDS-PAGE electrophoresis of purified RpfF protein. (b) RpfF reaction with acyl-ACPs, showing that RpfF cleaves acyl-ACP thioester bonds to release holo-ACP (lanes 2, 4, 6, 8); lane 9 contains only holo-ACP as a control. C8:0-ACP, octanoyl-ACP; C10:0-ACP, decanoyl-ACP; C12:0-ACP, dodecanoyl-ACP; C14:0-ACP, tetradencanoyl-ACP. (c) RpfF reaction with 3-hydroxydodecanoyl-ACP (3-OH-C12:0-ACP). Acyl-ACPs were first prepared as described in Materials and Methods. (d) RpfF reactions with 3-hydroxyacyl-ACPs. The cis-2-acyl-ACPs formed are indicated by arrows (lanes 2, 5, 7). Acyl-ACPs were prepared as described in Materials and Methods. (e) HPLC analysis of DSF extracts in the reaction mixture containing RpfF and 3-OH-C12:0-ACP (lane 5). LC-MS analysis showed that the active product in HPLC elute is BDSF.

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