Identification and Molecular Cloning of a Unique Hyaluronan Synthase from Pasteurella multocida

• DOI: 10.1074/jbc.273.14.8454
• Publication Date: 2002-07-26T15:04:43Z
• Authors: Paul L. DeAngelis; Wei Jing; Richard R. Drake; Ann Mary Achyuthan
• Keywords: Hyaluronan synthase; Cloning (programming); Identification

Type A Pasteurella multocida, a prevalent animal pathogen, employs hyaluronan [HA] polysaccharide capsule to avoid host defenses. We utilized transposon insertional mutagenesis identify the P. multocida HA synthase, enzyme that polymerizes HA. DNA fragment from wild-type genomic library could direct production in vivo Escherichia coli, bacterium normally does not produce Analysis of truncated plasmids derived original clone indicated an open reading frame encoding 972-residue protein was responsible for polymerization. This identification confirmed by expression cloning E. coli; we observed formation and detected activity membrane preparations vitro. The polypeptide size verified photoaffinity labeling native synthase with azido-UDP sugar analogs. Overall, sequence is very similar other known synthases streptococci, PBCV-1 virus, or vertebrates. Instead, portion central region new more homologous amino termini bacterial glycosyltransferases different capsular polysaccharides lipopolysaccharides. In summary, have discovered unique differs predicted topology enzymes.