Molecular genetic analysis of the nagH gene encoding a hyaluronidase of Clostridium perfringens.
A recombinant lambda phage was identified in a Clostridium perfringens genomic library by means of its ability to hydrolyse the fluorescent substrate 4-methyl-umbelliferyl-beta-D-glucosaminide, isolated and shown to encode an endo-beta-N-acetylglucosaminidase. This enzyme, NagH, is also known as hyaluronidase, or Mu toxin, a putative virulence factor which is likely to act on connective tissue during gas gangrene. Nucleotide sequence analysis allowed the primary structure to be deduced and showed hyaluronidase to be a large exported protein of 114,392 Daltons and an enzyme of this size, endowed with the corresponding activities, was partially purified from C. perfringens. Hyaluronidase seems to be organised into two domains, an N-terminal region comprising 700 amino acids bearing the active site and a 300-residue C-terminal segment, containing three copies of an extended motif. Two other reading frames, linked to nagH, also appear to encode proteins with sugar-binding motifs.
Canard
B
B
Unité de Génétique Moléculaire Bactérienne, Institut Pasteur, Paris, France.
Garnier
T
T
Saint-Joanis
B
B
Cole
S T
ST
D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D003016:Clostridium perfringens / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity; D004269:DNA, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D004926:Escherichia coli / Q000235:genetics; D005098:Exotoxins / Q000235:genetics; D005798:Genes, Bacterial; D006821:Hyaluronoglucosaminidase / Q000235:genetics*; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011993:Recombinant Fusion Proteins / Q000032:analysis; D015183:Restriction Mapping; D014774:Virulence / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005098:Exotoxins; D011993:Recombinant Fusion Proteins; C025158:4-methylumbelliferyl 2-acetamido-2-deoxy-beta-D-glucopyranoside; D006923:Hymecromone; D006821:Hyaluronoglucosaminidase; D000117:Acetylglucosamine