The essential protein for bacterial flagella formation FlgJ functions as a β-N-acetylglucosaminidase

Francesca A. Herlihey, Patrick Moynihan, Anthony Clarke

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36 Citations (Scopus)


The flagellum is a major virulence factor of motile pathogenic bacteria. This structure requires more than 50 proteins for its biogenesis and function, one of which is FlgJ. Homologs of FlgJ produced by the β- and γ-proteobacteria, such as Salmonella enterica, Vibrio spp., and both Sphingomonas sp. and Pseudomonas spp. are bifunctional, possessing an N-terminal domain responsible for proper rod assembly and a C-terminal domain possessing peptidoglycan lytic activity. Despite the amount of research conducted on FlgJ from these and other bacteria over the past 15 years, no biochemical analysis had been conducted on any FlgJ and consequently confusion exists as to whether the enzyme is a peptidoglycan hydrolase or a lytic transglycosylase. In this study, we present the development of a novel assay for glycoside lytic enzymes and its use to provide the first enzymatic characterization of the lytic domain of FlgJ from S. enterica as the model enzyme. Surprisingly, FlgJ functions as neither a muramidase nor a lytic transglycosylases but rather as a β-N-acetylglucosaminidase. As such, FlgJ represents the first autolysin with this activity to be characterized from a Gram-negative bacterium. At its optimal pH of 4.0, the Michaelis-Menten parameters of Km and kcat for FlgJ from S. enterica were determined to be 0.64 ± 0.18 mg ml-1 and 0.13 ± 0.016 s-1, respectively, using purified PG as substrate. Its catalytic residues were identified as Glu184 and Glu223.
Original languageEnglish
Pages (from-to)31029-31042
Number of pages14
JournalJournal of Biological Chemistry
Issue number45
Publication statusPublished - 23 Sept 2014


  • Acetylglucosaminidase
  • Amino Acid Sequence
  • Bacterial Proteins
  • Catalysis
  • Flagella
  • Glycolysis
  • Hydrogen-Ion Concentration
  • Kinetics
  • Mass Spectrometry
  • Molecular Sequence Data
  • N-Acetylmuramoyl-L-alanine Amidase
  • Open Reading Frames
  • Phylogeny
  • Plasmids
  • Protein Engineering
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Salmonella enterica
  • Sequence Homology, Amino Acid
  • Journal Article
  • Research Support, Non-U.S. Gov't
  • Bacterial Pathogenesis
  • Cell Wall
  • Enzyme Catalysis
  • Peptidoglycan
  • Virulence Factor
  • N-Acetylglycosaminidase
  • Autolysin
  • Lytic Transglycosylase
  • Muramidase


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