N- and O-linked glycosylation of total plasma glycoproteins in galactosemia

• Type of Material: Article
• Authors: Ying Liu, Emory University; Baoyun Xia, Emory University; Tyler J. Gleason, Emory University; Uriel Castaneda, Emory University; Miao He, Emory University; Gerard T. Berry, Children’s Hospital Boston; Judith Fridovich-Keil, Emory University
• Language: English
• Date: 2012-08-01
• Publisher: Elsevier: 12 months
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2012 Elsevier Inc. Published by Elsevier Inc. All rights reserved.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.ymgme.2012.05.025
• Title of Journal or Parent Work: Molecular Genetics and Metabolism
• ISSN: 1096-7192
• Volume: 106
• Issue: 4
• Start Page: 442
• End Page: 454
• Grant/Funding Information: The work was supported in part by grant NIH DK059904 (to JLFK) and in part by funds from the Manton Center for Orphan Disease Research (to GTB) and the Galactosemia Foundation (to GTB).; MH was supported in part by grant U54HD061939 from the Sterol and Isoprenoid Diseases (STAIR) consortium, a part of the NIH Rare Diseases clinical Research Network (RDCRN), funded by the NICHD and the NIH Office of Rare Diseases Research (ORDR).
• Keywords: HUMAN GALACTOSE-1-PHOSPHATE URIDYLYLTRANSFERASE; LC-MS/MS; Endocrinology & Metabolism; Genetics & Heredity; Medicine, Research & Experimental; GLYCANS; MASS-SPECTROMETRY; Plasma; Classic galactosemia; Science & Technology; Life Sciences & Biomedicine; DEFECTIVE GALACTOSYLATION; O-glycome; N-glycome; URIDYLTRANSFERASE; CLASSICAL GALACTOSEMIA; MALDI-TOF; OVARIAN-FUNCTION; Research & Experimental Medicine; ANTI-MULLERIAN HORMONE; SINGLE-FAMILY; SERUM TRANSFERRIN
• Research Categories: Biology, Genetics

Abstract

Classic galactosemia is a potentially lethal metabolic disorder that results from profound impairment of the enzyme galactose-1-phosphate uridylyltransferase (GALT); despite decades of research, the underlying mechanism of pathophysiology remains unclear. Previous studies of plasma and tissue samples from patients with classic galactosemia have revealed defects of protein and lipid glycosylation, however, the underlying bases for these defects and their clinical significance, if any, has remained unclear. As a step toward addressing these questions we c haracterized both the N- and O-linked glycomes of plasma proteins from neonates, infants, children, and adults with galactosemia using mass spectrometry and asked (1) whether similar or disparate defects exist for N-linked and O-linked modifications, (2) what factors correlate with the severity of these defects in different patients, and perhaps most important, (3) whether there is any apparent relationship between chronic glycosylation defects and long-term outcome in patients. We found that some but not all of the galactosemic neonates tested exhibited abnormal N- and O-linked glycosylation of plasma proteins. The types of abnormalities seen were similar between N- and O-linked moieties, but the extent of the defects varied between patients. Age, gender, GALT genotype, and predicted residual GALT activity all failed to explain the extent of the glycosylation defect in the samples studied. Dietary galactose restriction markedly normalized both the N- and O-linked glycosylation patterns for all infants tested; however, any remaining glycosylation defects evident in the plasma of older children or adults on galactose-restricted diets showed no correlation with clinical outcome. These data cannot rule out the possibility that subtle or localized glycosylation defects, not detectable by our methods or not reflected in plasma, may contribute to acute or long-term outcome severity.