The modular organization of multifunctional peptide synthetases

Joachim Vater, Torsten Stein, Dirk Vollenbroich, Volker Kruft, Brigitte Wittmann-Liebold, Peter Franke, Li Liu, Peter Zuber

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Gramicidin S synthetase 2 from B. brevis was affinity labeled at its valine thiolation center with the thiol reagent N-[3H]ethylmaleimide. From a tryptic digest of the enzyme-inhibitor complex a radioactive fragment was isolated in pure form by two reversed-phase HPLC steps. It was identified by liquid-phase N-terminal sequencing in combination with electrospray mass spectrometry (ESI-MS) as a hexadecapeptide containing the thiolation motif LGG(H/D)S(L/I). By ESI-MS it was demonstrated that a 4'-phosphopantetheine cofactor was attached to this fragment at its reactive serine. These results are consistent with the 'Multiple Carrier Model' of nonribosomal peptide biosynthesis. Site-specific mutagenesis has been performed in thiolation, elongation, and epimerization motifs of some of the modules of surfactin synthetase from B. subtilis to clarify the function of prominent conserved amino acid residues in the intermediate steps of peptide biosynthesis. The modular structure of multifunctional peptide synthetases is discussed.

Original languageEnglish (US)
Pages (from-to)557-564
Number of pages8
JournalJournal of Protein Chemistry
Issue number5
StatePublished - 1997
Externally publishedYes


  • Active site mutagenesis
  • Electrospray mass spectrometry
  • Modular structure
  • Multiple 4'-phosphopantetheine cofactors
  • Peptide synthetases
  • Thioester binding site

ASJC Scopus subject areas

  • Biochemistry


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