title: Comparison of Follicle-Stimulating Hormone Glycosylation Microheterogenity by Quantitative Negative Mode Nano-Electrospray Mass Spectrometry of Peptide-N Glycanase-Released Oligosaccharides.
abstract: Glycans from six highly purified hFSH preparations were released by peptide-N-glycanase digestion and analyzed by negative mode nano-ESI mass spectrometry before and after neuraminidase digestion. Pituitary glycan structures were mainly high-mannose, di-, tri-, and tetra-antennary, and their abundance largely paralleled that reported by other investigators using different approaches. For most of the FSH preparations, the differences in glycosylation appeared to be restricted to relative abundances of the major glycan families, as defined by their neutral core oligosaccharide structures. Qualitative differences between glycan populations were largely relegated to those species that were lowest in abundance. Significant qualitative differences were noted in two cases. Recombinant GH3-hFSH triantennary glycans appeared to have the third antenna exclusively on the mannose6-branch, in contrast to all pituitary and urinary hFSH triantennary glycans, in which this antenna was exclusively attached to the mannose3-branch. The hypo-glycosylated hFSH preparation isolated from purified hLH was decorated with high mannose glycans that accounted for over 40% of the total in this population. As this preparation was found to be consistently 20-fold more active than hFSH24in FSH receptor-binding assays, it appears that both macroheterogeneity and microheterogeneity in FSH preparations need to be taken into account.
authors.affiliation: Department of Biological Sciences, Wichita State University, Wichita, KS 67260.
authors.forename: George R
authors.initials: GR
authors.lastname: Bousfield
authors.affiliation: Department of Biological Sciences, Wichita State University, Wichita, KS 67260.
authors.forename: Vladimir Y
authors.initials: VY
authors.lastname: Butnev
authors.affiliation: Department of Biological Sciences, Wichita State University, Wichita, KS 67260.
authors.forename: William K
authors.initials: WK
authors.lastname: White
authors.affiliation: Molecular Graphics and Modeling Laboratory, University of Kansas, Lawrence, KS 66045.
authors.forename: Aaron Smalter
authors.initials: AS
authors.lastname: Hall
authors.affiliation: Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Harvey
keywords: Follicle-stimulating hormone; mass spectrometry; oligosaccharides