title: Identification of new apolipoprotein-CIII glycoforms with ultrahigh resolution MALDI-FTICR mass spectrometry of human sera.
abstract: Apolipoprotein-CIII (apoCIII) is an abundant blood glycoprotein associated with lipoprotein particles. Three different glycoforms have been described, all containing a mucin-type core-1 O-glycosylation with either zero, one or two sialic acids. Changes in the relative abundance of these glycoforms have been observed in a variety of different pathologies. In this study, ultrahigh resolution 15T MALDI Fourier transform ion cyclotron resonance (FTICR) MS was used to analyze apoCIII isoforms in serum protein profiles. For this purpose, serum proteins were purified using both a fully automated RPC18-based magnetic bead method and an RPC4 cartridge-based solid phase extraction method. Six new apoCIII isoforms were identified with low-ppm mass measurement errors and ultrahigh precision. These were characterized by more complex glycan moieties that are fucosylated instead of sialylated. To confirm the glycan moiety and localize the glycosylation site, top-down ESI-FTICR-MS/MS and bottom-up LC-ion trap MS/MS were used. A large variation in the presence and abundance of the fucosylated isoforms was found in a set of 96 serum samples. These findings of fucosylated apolipoprotein-CIII isoforms warrant further research to elucidate the implications these glycoforms may have for the plethora of studies where alterations in apoCIII have been linked to the development of many different pathologies.
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center (LUMC), Albinusdreef 2, 2300 RC, Leiden, The Netherlands. s.nicolardi@lumc.nl
authors.forename: Simone
authors.initials: S
authors.lastname: Nicolardi
authors.forename: Yuri E M
authors.initials: YE
authors.lastname: van der Burgt
authors.forename: Irina
authors.initials: I
authors.lastname: Dragan
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hensbergen
authors.forename: André M
authors.initials: AM
authors.lastname: Deelder
mesh_terms: D000595:Amino Acid Sequence; D053305:Apolipoprotein C-III / Q000097:blood* / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D005583:Fourier Analysis; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D053305:Apolipoprotein C-III; D001798:Blood Proteins