title: The Actinobacillus pleuropneumoniae HMW1C-like glycosyltransferase mediates N-linked glycosylation of the Haemophilus influenzae HMW1 adhesin.
abstract: The Haemophilus influenzae HMW1 adhesin is an important virulence exoprotein that is secreted via the two-partner secretion pathway and is glycosylated at multiple asparagine residues in consensus N-linked sequons. Unlike the heavily branched glycans found in eukaryotic N-linked glycoproteins, the modifying glycan structures in HMW1 are mono-hexoses or di-hexoses. Recent work demonstrated that the H. influenzae HMW1C protein is the glycosyltransferase responsible for transferring glucose and galactose to the acceptor sites of HMW1. An Actinobacillus pleuropneumoniae protein designated ApHMW1C shares high-level homology with HMW1C and has been assigned to the GT41 family, which otherwise contains only O-glycosyltransferases. In this study, we demonstrated that ApHMW1C has N-glycosyltransferase activity and is able to transfer glucose and galactose to known asparagine sites in HMW1. In addition, we found that ApHMW1C is able to complement a deficiency of HMW1C and mediate HMW1 glycosylation and adhesive activity in whole bacteria. Initial structure-function studies suggested that ApHMW1C consists of two domains, including a 15-kDa N-terminal domain and a 55-kDa C-terminal domain harboring glycosyltransferase activity. These findings suggest a new subfamily of HMW1C-like glycosyltransferases distinct from other GT41 family O-glycosyltransferases.
authors.affiliation: Department of Biology and Biochemistry, University of Houston, Houston, Texas, United States of America.
authors.forename: Kyoung-Jae
authors.initials: KJ
authors.lastname: Choi
authors.forename: Susan
authors.initials: S
authors.lastname: Grass
authors.forename: Seonghee
authors.initials: S
authors.lastname: Paek
authors.forename: Joseph W
authors.initials: JW
authors.lastname: St Geme
authors.forename: Hye-Jeong
authors.initials: HJ
authors.lastname: Yeo
mesh_terms: D016977:Actinobacillus pleuropneumoniae / Q000378:metabolism*; D018829:Adhesins, Bacterial / Q000737:chemistry*; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D002384:Catalysis; D019295:Computational Biology / Q000379:methods; D004847:Epithelial Cells / Q000382:microbiology; D006031:Glycosylation; D016695:Glycosyltransferases / Q000737:chemistry*; D006193:Haemophilus influenzae / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid
chemical_list: D018829:Adhesins, Bacterial; C117431:HMW1 protein, bacteria; D016695:Glycosyltransferases