title: A new enzyme superfamily - the phosphopantetheinyl transferases.
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 o195); ACPS and EntD are specific for the activation of fatty acid synthase and enterobactin synthetase, respectively. The apo-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.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA. walsh@walsh.med.harvard.edu
authors.forename: R H
authors.initials: RH
authors.lastname: Lambalot
authors.forename: A M
authors.initials: AM
authors.lastname: Gehring
authors.forename: R S
authors.initials: RS
authors.lastname: Flugel
authors.forename: P
authors.initials: P
authors.lastname: Zuber
authors.forename: M
authors.initials: M
authors.lastname: LaCelle
authors.forename: M A
authors.initials: MA
authors.lastname: Marahiel
authors.forename: R
authors.initials: R
authors.lastname: Reid
authors.forename: C
authors.initials: C
authors.lastname: Khosla
authors.forename: C T
authors.initials: CT
authors.lastname: Walsh
mesh_terms: D000595:Amino Acid Sequence; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014166:Transferases / Q000737:chemistry* / Q000378:metabolism; D014633:Valine / Q000378:metabolism
chemical_list: D014166:Transferases; D014633:Valine