Characterization of the formation of the pyrrole moiety during clorobiocin and coumermycin A1 biosynthesis

Sylvie Garneau, Pieter C. Dorrestein, Neil L. Kelleher, Christopher T. Walsh

Research output: Contribution to journalArticlepeer-review

74 Scopus citations


The aminocoumarin antibiotics clorobiocin and coumermycin A1 target the B subunit of DNA gyrase by presentation of the 5-methyl-pyrrolyl-2- carboxy ester moiety in the ATP-binding site of the enzyme. The pyrrolyl pharmacophore is derived by a four electron oxidation of a prolyl unit while tethered in phosphopantetheinyl thioester linkage to a peptidyl carrier protein (PCP) subunit. L-Proline is selected and activated as L-prolyl-AMP by adenylation domain enzymes (CloN4 and CouN4) and then installed as the thioester on the holo form of the PCP proteins CloN5 and CouN5. Enzymatic oxidation of the prolyl-S-PCP by the flavoprotein dehydrogenase CloN3 can be followed by rapid quench and subsequent electrospray ionization - Fourier transform mass spectrometry analysis of the acyl-S-protein substrate/ product mixture to establish that a two-electron oxidized pyrrolinyl-S-enzyme transiently accumulates on the way to the four-electron oxidized, heteroaromatic pyrrolyl-2-carboxy-S-PCP acyl enzyme product.

Original languageEnglish
Pages (from-to)2770-2780
Number of pages11
Issue number8
StatePublished - Mar 1 2005

ASJC Scopus subject areas

  • Biochemistry


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