title: N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M.
abstract: In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213.
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, Cagliari, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Manconi
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, Cagliari, Italy.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Cabras
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, Cagliari, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Sanna
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, Cagliari, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Piras
authors.affiliation: Dipartimento di Scienze della Vita e dell'Ambiente, Università di Cagliari, Cagliari, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Liori
authors.affiliation: Dipartimento di Scienze Chirurgiche, Università di Cagliari, Cagliari, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Pisano
authors.affiliation: Istituto di Biochimica e Biochimica Clinica, Università Cattolica, Roma, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Iavarone
authors.affiliation: Istituto di Biochimica e Biochimica Clinica, Università Cattolica, Roma, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Vincenzoni
authors.affiliation: Istituto di Clinica Odontostomatologica, Facoltà di Medicina, Università Cattolica, Roma, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Cordaro
authors.affiliation: Dipartimento di Scienze Chirurgiche, Università di Cagliari, Cagliari, Italy.
authors.forename: Gavino
authors.initials: G
authors.lastname: Faa
authors.affiliation: Istituto di Biochimica e Biochimica Clinica, Università Cattolica, Roma, Italy.|Istituto di Chimica del Riconoscimento Molecolare - CNR, Roma, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Castagnola
authors.affiliation: Istituto di Chimica del Riconoscimento Molecolare - CNR, Roma, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Messana
mesh_terms: D002851:Chromatography, High Pressure Liquid; D006031:Glycosylation; D006801:Humans; D011134:Polysaccharides / Q000737:chemistry*; D055273:Salivary Proline-Rich Proteins / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D011134:Polysaccharides; D055273:Salivary Proline-Rich Proteins
keywords: Mass Spectrometry; N-Deglycosylation; Saliva; Site-specific glycosylation