Title
HPLC analysis of discrete haptoglobin isoform N-linked oligosaccharides following 2D-PAGE isolation
Document Type
Journal Article
RIS ID
39021
Abstract
Glycosylation is a common but variable modification that regulates glycoprotein structure and function. We combined small format 2D-PAGE with HPLC to analyse discrete human haptoglobin isoform N-glycans. Seven major and several minor haptoglobin isoforms were detected by 2D-PAGE. N-Glycans released from Coomassie-stained gel spots using PNGase were labeled at their reducing termini with 2-aminobenzamide. HPLC analysis of selected major isoform N-glycans indicated that sialic acid composition determined their Separation by isoelectric focussing. N-Glycans from two doublets of quantitatively minor isoforms were also analysed. Although separation of each pair of doublets was influenced by sialylation, individual spots within each doublet contained identical N-glycans. Thus, heterogeneity in minor haptoglobin isoforms was due to modifications distinct from N-glycan structure. These studies describe a simple method for analysing low abundance protein N-glycans and provide details of discrete haptoglobin isoform N-glycan structures which will be useful in proteomic analysis of human plasma samples. (c) 2006 Elsevier Inc. All rights reserved.
This record is in the process of being updated. Please contact us for more information.
Publication Details
He, Z., Aristoteli, L. P., Kritharides, L. & Garner, B. (2006). HPLC analysis of discrete haptoglobin isoform N-linked oligosaccharides following 2D-PAGE isolation. Biochemical and Biophysical Research Communications, 343 (2), 496-503.