FIGURE 5.

Vc0213 is a myristoyl (C14:0) acyltransferase and utilizes C14:0-CoA as its preferred acyl donor. A, MKV15 (DE3) membranes (0.01 mg/ml) expressing either Bp3073, Hi1527, or Vc0213 were assayed with Kdo₂-[4′-³²P]lipid IV_A and individual acyl-ACPs (C12:0, C14:0, C16:0, or C18:0) for 30 min. The percentage of conversion was calculated based on the amount of Kdo₂-[4′-³²P]lipid IV_A converted to the acylated form. B, membranes expressing Vc0213, pBp3073, or pHi1527 (0.01 mg/ml) were assayed for 15 min. with either C14:0-ACP or C14:0-CoA using phosphorylated Kdo-[4′-³²P]lipid IV_A as the lipid acceptor. For assays containing E. coli LpxL, the concentration of membranes was 0.001 mg/ml and either C12:0-ACP or C12:0-CoA and Kdo₂-[4′-³²P]lipid IV_A as the lipid acceptor.