title: Application of multidimensional affinity high-performance liquid chromatography and electrospray ionization liquid chromatography-mass spectrometry to the characterization of glycosylation in single-chain plasminogen activator. Initial results.
abstract: Preliminary results are presented using a combination of affinity chromatography, reversed-phase HPLC and electrospray ionization mass spectrometry to produced peptide maps for N-linked, O-linked and non-glycosylated peptides from an endoproteinase LysC digest of DSPA alpha 1, a recombinant DNA derived glycoprotein. Although the system was used to identify a number of major N-linked structures, notably complex biantennary structures attached to asparagine 362, no O-linked glycopeptides from the possible 4 attachment sites were identified. The system did, however, demonstrate the feasibility of the approach and the applicability of the instrumental system.
authors.affiliation: Biomeasurements Group, Hewlett-Packard Laboratories, Palo Alto, CA 94304, USA.
authors.forename: A
authors.initials: A
authors.lastname: Apffel
authors.forename: J A
authors.initials: JA
authors.lastname: Chakel
authors.forename: W S
authors.initials: WS
authors.lastname: Hancock
authors.forename: C
authors.initials: C
authors.lastname: Souders
authors.forename: T
authors.initials: T
authors.lastname: M'Timkulu
authors.forename: E
authors.initials: E
authors.lastname: Pungor
mesh_terms: D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D003208:Concanavalin A / Q000737:chemistry; D006031:Glycosylation; D006241:Haptens; D037102:Lectins / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010960:Plasminogen Activators / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity
chemical_list: D006241:Haptens; D037102:Lectins; D003208:Concanavalin A; D010960:Plasminogen Activators