title: Structure and bacterial receptor activity of a human salivary proline-rich glycoprotein.
abstract: Using an overlay technique, we previously showed that the Gram-negative periodontal pathogen Fusobacterium nucleatum binds to a glycoprotein of Mr 89,000 (Prakobphol, A., Murray, P., and Fischer, S.J. (1987) Anal. Biochem. 164, 5-11) in the parotid saliva of some individuals. We now show that deglycosylation of the purified glycoprotein results in loss of receptor activity. Amino acid analysis of the protein core showed predominantly proline, glycine, and glutamic acid/glutamine, a characteristic of proline-rich glycoproteins (PRG). The amino terminus contained repeating sequences of Ser-Gln-Gly-Pro-Pro-Pro-Arg-Pro-Gly-Lys-Pro-Glu-Gly-Pro-Pro-Pro- Gln-Gly that had significant compositional and sequence homology to that encoded by exon 3 of the PRB3 gene. We analyzed the PRG oligosaccharides by a combination of mass spectrometry techniques and nuclear magnetic resonance spectroscopy. Twenty-seven highly fucosylated structures were identified. The most abundant was as follows (where Fuc is fucose). (formula; see text) To understand the structural basis of F. nucleatum binding, we screened glycolipids and neoglycolipids carrying carbohydrate structures related to those of the PRG for receptor activity; components with unsubstituted terminal lactosamine residues best supported adherence. Neoglycolipids constructed from PRG oligosaccharides were also receptors. Treatment with beta-galactosidase, but not alpha-fucosidase, abolished binding, suggesting that unsubstituted lactosamine units, including the 6-antenna of the major oligosaccharide, mediate F. nucleatum adherence.
authors.affiliation: Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco 94143.
authors.forename: B L
authors.initials: BL
authors.lastname: Gillece-Castro
authors.forename: A
authors.initials: A
authors.lastname: Prakobphol
authors.forename: A L
authors.initials: AL
authors.lastname: Burlingame
authors.forename: H
authors.initials: H
authors.lastname: Leffler
authors.forename: S J
authors.initials: SJ
authors.lastname: Fisher
mesh_terms: D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D005673:Fusobacterium / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D055232:Proline-Rich Protein Domains; D012471:Salivary Proteins and Peptides / Q000737:chemistry* / Q000378:metabolism
chemical_list: D010455:Peptides; D012471:Salivary Proteins and Peptides