We experimentally identified the activities of six predicted heptosyltransferases in Actinobacillus pleuropneumoniae genome serotype 5b strain L20 and serotype 3 strain JL03. The initial identification was based on a bioinformatic analysis of the amino acid similarity between these putative heptosyltrasferases with others of known function from enteric bacteria and Aeromonas. The putative functions of all the Actinobacillus pleuropneumoniae heptosyltrasferases were determined by using surrogate LPS acceptor molecules from well-defined A. hydrophyla AH-3 and A. salmonicida A450 mutants. Our results show that heptosyltransferases APL_0981 and APJL_1001 are responsible for the transfer of the terminal outer core D-glycero-D-manno-heptose (D,D-Hep) residue although they are not currently included in the CAZY glycosyltransferase 9 family. The WahF heptosyltransferase group signature sequence [S(T/S)(GA)XXH] differs from the heptosyltransferases consensus signature sequence [D(TS)(GA)XXH], because of the substitution of D(261) for S(261), being unique.
serotype, heptosyltransferase, genome, Actinobacillus pleuropneumoniae
NCBI PubMed ID: 23383222Publication DOI: 10.1371/journal.pone.0055546Journal NLM ID: 101285081Publisher: San Francisco, CA: Public Library of Science
Correspondence: jtomas@ub.edu
Institutions: Departamento de Microbiologia, Facultad de Biologia, Universidad de Barcelona, Barcelona, Spain
Methods: SDS-PAGE, DNA techniques, ESI-MS, mild acid hydrolysis, genetic methods, bioinformatic analysis
Found