title: Bottom-Up Elucidation of Glycosidic Bond Stereochemistry.
abstract: The lack of robust, high-throughput, and sensitive analytical strategies that can conclusively map the structure of glycans has significantly hampered progress in fundamental and applied aspects of glycoscience. Resolution of the anomeric α/β glycan linkage within oligosaccharides remains a particular challenge. Here, we show that "memory" of anomeric configuration is retained following gas-phase glycosidic bond fragmentation during tandem mass spectrometry (MS2). These findings allow for integration of MS2with ion mobility spectrometry (IM-MS2) and lead to a strategy to distinguish α- and β-linkages within natural underivatized carbohydrates. We have applied this fragment-based hyphenated MS technology to oligosaccharide standards and to de novo sequencing of purified plant metabolite glycoconjugates, showing that the anomeric signature is also observable in fragments derived from larger glycans. The discovery of the unexpected anomeric memory effect is further supported by IR-MS action spectroscopy and ab initio calculations. Quantum mechanical calculations provide candidate geometries for the distinct anomeric fragment ions, in turn shedding light on gas-phase dissociation mechanisms of glycosidic linkages.
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester, M1 7DN, United Kingdom.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Gray
authors.affiliation: Institut Lumière Matière, UMR5306 Université Lyon 1-CNRS, Université de Lyon 69622 Villeurbanne Cedex, France.
authors.forename: Baptiste
authors.identifier: 0000-0002-7376-4154
authors.initials: B
authors.lastname: Schindler
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester, M1 7DN, United Kingdom.
authors.forename: Lukasz G
authors.initials: LG
authors.lastname: Migas
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences and Center for Synthetic Biology, University of Copenhagen , 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Martina
authors.initials: M
authors.lastname: Pičmanová
authors.affiliation: Institut Lumière Matière, UMR5306 Université Lyon 1-CNRS, Université de Lyon 69622 Villeurbanne Cedex, France.
authors.forename: Abdul R
authors.initials: AR
authors.lastname: Allouche
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester, M1 7DN, United Kingdom.
authors.forename: Anthony P
authors.initials: AP
authors.lastname: Green
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester, M1 7DN, United Kingdom.
authors.forename: Santanu
authors.initials: S
authors.lastname: Mandal
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences and Center for Synthetic Biology, University of Copenhagen , 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Mohammed S
authors.initials: MS
authors.lastname: Motawia
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences and Center for Synthetic Biology, University of Copenhagen , 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Raquel
authors.initials: R
authors.lastname: Sánchez-Pérez
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences and Center for Synthetic Biology, University of Copenhagen , 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Nanna
authors.initials: N
authors.lastname: Bjarnholt
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences and Center for Synthetic Biology, University of Copenhagen , 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Birger L
authors.initials: BL
authors.lastname: Møller
authors.affiliation: Radboud University , Institute for Molecules and Materials, FELIX Laboratory, Toernooiveld 7c, 6525ED Nijmegen, The Netherlands.
authors.forename: Anouk M
authors.initials: AM
authors.lastname: Rijs
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester, M1 7DN, United Kingdom.|Michael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester, M1 7DN, United Kingdom.
authors.forename: Perdita E
authors.identifier: 0000-0002-7720-586X
authors.initials: PE
authors.lastname: Barran
authors.affiliation: Institut Lumière Matière, UMR5306 Université Lyon 1-CNRS, Université de Lyon 69622 Villeurbanne Cedex, France.|Institut Universitaire de France IUF , 103 Boulevard St. Michel, 75005 Paris, France.
authors.forename: Isabelle
authors.identifier: 0000-0003-2994-3961
authors.initials: I
authors.lastname: Compagnon
authors.affiliation: Department of Biochemistry, Institute of Integrative Biology, University of Liverpool , Crown Street, Liverpool, L69 7ZB, United Kingdom.
authors.forename: Claire E
authors.initials: CE
authors.lastname: Eyers
authors.affiliation: School of Chemistry & Manchester Institute of Biotechnology, The University of Manchester , 131 Princess Street, Manchester, M1 7DN, United Kingdom.
authors.forename: Sabine L
authors.identifier: 0000-0003-3974-646X
authors.initials: SL
authors.lastname: Flitsch