title: Human gastric TFF2 peptide contains an N-linked fucosylated N,N'-diacetyllactosediamine (LacdiNAc) oligosaccharide.
abstract: In the human stomach, the peptide trefoil factor family 2 (TFF2) is secreted together with the mucin MUC6 by mucous neck cells (MNCs) and antral gland cells. TFF2 is strongly associated with the gastric mucus and promotes gastric restitution. Here, TFF2 was purified from the human corpus and antrum, respectively, by size-exclusion chromatography, and the N-linked glycan structure at N-15 of the mature peptide was determined. As a hallmark, the unusual monofucosylated N,N'-diacetylhexosediamine (tentatively assigned as GalNAcβ1 → 4GlcNAc, LacdiNAc) modification was detected as the terminal structure of a bi-antennary complex type N-glycan exhibiting also core fucosylation. Replicate analyses did not show microheterogeneities in the fraction of peptide-N-glycosidase F cleaved and permethylated N-glycans when analyzed by matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS). On the glycopeptide level, a minor glycan microheterogeneity was evident in liquid chromatography-electrospray ionization (ESI)-MS, demonstrating the presence of underfucosylated species. The tryptic TFF2 N-glycopeptide p34-39 (LSPHNR N-glycosylated with Fuc3Hex3HexNAc6) was identified by both ESI-tandem mass spectrometry and MALDI-post-source decay analysis. Lectin analyses with the Wisteria floribunda agglutinin indicated the potential presence of LacdiNAc terminating glycans and revealed minor differences between TFF2 from fundic units, i.e. MNCs, and antral units, i.e. antral gland cells. Strikingly, on the level of the primary structure, there was no indication that the formation of the proposed LacdiNAc structure is cis-controlled by a peptidic determinant related to the published sequences.
authors.affiliation: Institute of Biochemistry II, Medical Faculty, and Central Bioanalytics, Center for Molecular Medicine Cologne, University Köln, Germany.
authors.forename: Franz-Georg
authors.initials: FG
authors.lastname: Hanisch
authors.forename: Hubert
authors.initials: H
authors.lastname: Ragge
authors.forename: Thomas
authors.initials: T
authors.lastname: Kalinski
authors.forename: Frank
authors.initials: F
authors.lastname: Meyer
authors.forename: Hubert
authors.initials: H
authors.lastname: Kalbacher
authors.forename: Werner
authors.initials: W
authors.lastname: Hoffmann
mesh_terms: D000595:Amino Acid Sequence; D006801:Humans; D007785:Lactose / Q000031:analogs & derivatives* / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry / Q000378:metabolism; D034781:Receptors, N-Acetylglucosamine / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000071164:Trefoil Factor-2
chemical_list: D009844:Oligosaccharides; D010455:Peptides; D037121:Plant Lectins; D034781:Receptors, N-Acetylglucosamine; C000604007:TFF2 protein, human; D000071164:Trefoil Factor-2; C040129:wisteria lectin; C093701:N-acetylgalactosaminyl-1-4-N-acetylglucosamine; D007785:Lactose