A general strategy to enzymatically label acyl carrier proteins (ACPs) of polyketide synthases has been developed. Incorporation of a chloromethyl ketone or vinyl ketone moiety into polyketide chain elongation intermediate mimics allows for the synthesis of CoA adducts. These CoA adducts undergo enzymatic reaction with Sfp, a phosphopantetheinyl transferase, to afford labeled CurB carrier proteins.
Bibliographical noteFunding Information:
The authors thank Liangcai Gu and David Sherman for plasmid pET28b::CurB encoding CurB, K. R. Noon (Biomedical Mass Spectrometry Facility, University of Michigan) for assistance with LC–MS, and Jamie Razelun for preparation of CurB. Financial support from NIH DK-42303 (to J.L.S.) and NIH GM081544-01 (to J.L.S. and R.A.F.) is gratefully acknowledged.
- Acyl carrier proteins