A new enzyme superfamily - The phosphopantetheinyl transferases

Ralph H. Lambalot, Amy M. Gehring, Roger S. Flugel, Peter Zuber, Michael LaCelle, Mohamed A. Marahiel, Ralph Reid, Chaitan Khosla, Christopher T. Walsh

Research output: Contribution to journalArticle

646 Citations (Scopus)

Abstract

Background: All polyketide synthases, fatty acid synthases, and non- ribosomal peptide synthetases require posttranslational modification of their constituent acyl carrier protein domain(s) to become catalytically active. The inactive apoproteins are converted to their active holo-forms by posttranslational transfer of the 4'-phosphopantetheinyl (P-pant) moiety of coenzyme A to the sidechain hydroxyl of a conserved serine residue in each acyl carrier protein domain. The first P-pant transferase to be cloned and characterized was the recently reported Escherichia coli enzyme ACPS, responsible for apo to holo conversion of fatty acid synthase. Surprisingly, initial searches of sequence databases did not reveal any proteins with significant peptide sequence similarity with ACPS. Results: Through refinement of sequence alignments that indicated low level similarity with the ACPS peptide sequence, we identified two consensus motifs shared among several potential ACPS homologs. This has led to the identification of a large family of proteins having 12-22% similarity with ACPS, which are putative P-pant transferases. Three of these proteins, E. coli EntD and o195, and B. subtilis Sfp, have been overproduced, purified and found to have P- pant transferase activity, confirming that the observed low level of sequence homology correctly predicted catalytic function. Three P-pant transferases are now known to be present in E. coli (ACPS, EntD and o 195); ACPS and EntD are specific for the activation of fatty acid synthase and enterobactin synthetase, respectively. The ape-protein substrate for o195 has not yet been identified. Sfp is responsible for the activation of the surfactin synthetase. Conclusions: The specificity of ACPS and EntD for distinct P- pant-requiring enzymes suggests that each P-pant-requiring synthase has its own partner enzyme responsible for apo to holo activation of its acyl carrier domains. This is the first direct evidence that in organisms containing multiple P-pant-requiring pathways, each pathway has its own posttranslational modifying activity.

Original languageEnglish (US)
Pages (from-to)923-936
Number of pages14
JournalChemistry and Biology
Volume3
Issue number11
StatePublished - Nov 1996
Externally publishedYes

Fingerprint

Fatty Acid Synthases
Acyl Carrier Protein
Chemical activation
Escherichia coli
Enzymes
Peptide Synthases
Polyketide Synthases
Peptides
Proteins
Apoproteins
Escherichia coli Proteins
Coenzyme A
Transferases
Hydroxyl Radical
Serine
Sequence Alignment
Hominidae
Post Translational Protein Processing
Sequence Homology
Databases

Keywords

  • ACP
  • acyl carrier protein
  • biosynthesis
  • non-ribosomal peptide synthetase
  • phosphopantetheine

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Lambalot, R. H., Gehring, A. M., Flugel, R. S., Zuber, P., LaCelle, M., Marahiel, M. A., ... Walsh, C. T. (1996). A new enzyme superfamily - The phosphopantetheinyl transferases. Chemistry and Biology, 3(11), 923-936.

A new enzyme superfamily - The phosphopantetheinyl transferases. / Lambalot, Ralph H.; Gehring, Amy M.; Flugel, Roger S.; Zuber, Peter; LaCelle, Michael; Marahiel, Mohamed A.; Reid, Ralph; Khosla, Chaitan; Walsh, Christopher T.

In: Chemistry and Biology, Vol. 3, No. 11, 11.1996, p. 923-936.

Research output: Contribution to journalArticle

Lambalot, RH, Gehring, AM, Flugel, RS, Zuber, P, LaCelle, M, Marahiel, MA, Reid, R, Khosla, C & Walsh, CT 1996, 'A new enzyme superfamily - The phosphopantetheinyl transferases', Chemistry and Biology, vol. 3, no. 11, pp. 923-936.
Lambalot RH, Gehring AM, Flugel RS, Zuber P, LaCelle M, Marahiel MA et al. A new enzyme superfamily - The phosphopantetheinyl transferases. Chemistry and Biology. 1996 Nov;3(11):923-936.
Lambalot, Ralph H. ; Gehring, Amy M. ; Flugel, Roger S. ; Zuber, Peter ; LaCelle, Michael ; Marahiel, Mohamed A. ; Reid, Ralph ; Khosla, Chaitan ; Walsh, Christopher T. / A new enzyme superfamily - The phosphopantetheinyl transferases. In: Chemistry and Biology. 1996 ; Vol. 3, No. 11. pp. 923-936.
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