title: Determination of the Disulfide Structure of Murine Meteorin, a Neurotrophic Factor, by LC-MS and Electron Transfer Dissociation-High-Energy Collisional Dissociation Analysis of Proteolytic Fragments.
abstract: Meteorin and Cometin (Meteorin-like) are secreted proteins belonging to a newly discovered growth factor family. Both proteins play important roles in neural development and may have potential as therapeutic targets or agents. Meteorin and Cometin are homologues and contain ten evolutionarily conserved Cys residues across a wide variety of species. However, the status of the Cys residues has remained unknown. Here, we have successfully determined the disulfide structure for murine Meteorin by LC-MS analysis of fragments generated by trypsin plus endoprotease-Asp-N. For proteolytic fragments linked by more than one disulfide bond, we used electron transfer dissociation (ETD) to partially dissociate disulfide bonds followed by high-energy collisional dissociation (HCD) to determine disulfide linkages. Our analysis revealed that the ten Cys residues in murine Meteorin form five disulfide bonds with Cys7 (C1) linked to Cys28 (C2), Cys59 (C3) to Cys95 (C4), Cys148 (C5) to Cys219 (C8), Cys151 (C6) to Cys243 (C9), and Cys161 (C7) to Cys266 (C10). Since the ten Cys residues are highly conserved in Meteorin and Cometin, it is likely that the disulfide linkages are also conserved. This disulfide structure information should facilitate structure-function relationship studies on this new class of neurotrophic factors and also assist in evaluation of their therapeutic potentials.
authors.affiliation: Department of Protein Drug Discovery, Biogen Inc. , 225 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Dingyi
authors.identifier: 0000-0001-6631-2136
authors.initials: D
authors.lastname: Wen
authors.affiliation: Department of Protein Drug Discovery, Biogen Inc. , 225 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Department of Protein Drug Discovery, Biogen Inc. , 225 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Malgorzata M
authors.initials: MM
authors.lastname: Vecchi
authors.affiliation: Department of Protein Drug Discovery, Biogen Inc. , 225 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Bang Jian
authors.initials: BJ
authors.lastname: Gong
authors.affiliation: Department of Protein Drug Discovery, Biogen Inc. , 225 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Jana
authors.initials: J
authors.lastname: Dolnikova
authors.affiliation: Department of Protein Drug Discovery, Biogen Inc. , 225 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: R Blake
authors.initials: RB
authors.lastname: Pepinsky
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D004220:Disulfides / Q000032:analysis*; D004579:Electron Transport; D004735:Energy Transfer; D051379:Mice; D015394:Molecular Structure; D009414:Nerve Growth Factors / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D059748:Proteolysis; D053719:Tandem Mass Spectrometry
chemical_list: D004220:Disulfides; D009414:Nerve Growth Factors; D010446:Peptide Fragments