Posttranslational formation of formylglycine in prokaryotic sulfatases by modification of either cysteine or serine

J Biol Chem. 1998 Oct 2;273(40):25560-4. doi: 10.1074/jbc.273.40.25560.

Abstract

Eukaryotic sulfatases carry an alpha-formylglycine residue that is essential for activity and is located within the catalytic site. This formylglycine is generated by posttranslational modification of a conserved cysteine residue. The arylsulfatase gene of Pseudomonas aeruginosa also encodes a cysteine at the critical position. This protein could be expressed in active form in a sulfatase-deficient strain of P. aeruginosa, thereby restoring growth on aromatic sulfates as sole sulfur source, and in Escherichia coli. Analysis of the mature protein expressed in E. coli revealed the presence of formylglycine at the expected position, showing that the cysteine is also converted to formylglycine in a prokaryotic sulfatase. Substituting the relevant cysteine by a serine codon in the P. aeruginosa gene led to expression of inactive sulfatase protein, lacking the formylglycine. The machinery catalyzing the modification of the Pseudomonas sulfatase in E. coli therefore resembles the eukaryotic machinery, accepting cysteine but not serine as a modification substrate. By contrast, in the arylsulfatase of Klebsiella pneumoniae a formylglycine is found generated by modification of a serine residue. The expression of both the Klebsiella and the Pseudomonas sulfatases as active enzymes in E. coli suggests that two modification systems are present, or that a common modification system is modulated by a cofactor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / analogs & derivatives*
  • Alanine / biosynthesis
  • Arylsulfatases / chemistry*
  • Arylsulfatases / genetics
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Binding Sites / genetics
  • Borohydrides / metabolism
  • Cysteine / genetics
  • Glycine / analogs & derivatives*
  • Glycine / biosynthesis
  • Klebsiella pneumoniae / enzymology
  • Mutagenesis, Site-Directed
  • Peptide Fragments / chemistry
  • Prokaryotic Cells / enzymology*
  • Protein Processing, Post-Translational / genetics*
  • Pseudomonas aeruginosa / enzymology
  • Recombinant Proteins / chemistry
  • Sequence Analysis
  • Serine / genetics*
  • Sulfates / metabolism
  • Trypsin / metabolism

Substances

  • Bacterial Proteins
  • Borohydrides
  • Peptide Fragments
  • Recombinant Proteins
  • Sulfates
  • Serine
  • C(alpha)-formylglycine
  • sodium borohydride
  • Arylsulfatases
  • Trypsin
  • Cysteine
  • Alanine
  • Glycine