Differential Glycomics of Epithelial Membrane Glycoproteins from Urinary Exovesicles Reveals Shifts toward Complex-Type N-Glycosylation in Classical Galactosemia

作者：Staubach Simon; Schadewaldt Peter; Wendel Udo; Nohroudi Klaus; Hanisch Franz Georg^*

来源：Journal of Proteome Research, 2012, 11(2): 906-916.

DOI：10.1021/pr200711w

• classical galactosemia
• differential glycomics
• urinary exosomes
• membrane glycoprotein
• Tamm-Horsfall glycoprotein
• N-glycosylation
• MALDI mass spectrometry

A variety of genetic variations in the galactose-1-phosphate uridyltransferase (GALT) gene cause profound activity loss of the enzyme and acute toxic effects mediated by accumulating metabolic intermediates of galactose in newborns induced by dietary galactose. However, even on a severely galactose-restricted diet, patients develop serious long-term complications of the CNS and ovaries, which may result from damaging perturbations in cell biology caused by endogenously synthezised galactose. Under galactose stress, the cosubstrate of GALT, galactose-1-phosphate, accumulates and disturbs catabolic and anabolic pathways of the carbohydrate metabolism with potential effects on protein glycosylation and membrane localization of glycoprotein receptors, like the epidermal growth factor receptor. To address this issue in view of a cellular pathomechanism, we performed a differential semiquantitative N-glycomics study of membrane proteins. A suitable noninvasive cellular material derived from epithelial plasma membranes was found in urinary exovesicles and in the shed Tamm-Horsfall protein. By applying matrix-assisted laser ionization mass spectrometry on permethylated, PNGaseF released N-glycans, we demonstrate that GALT deficiency is associated with dramatic shifts from prevalent high-mannose-type glycans found in healthy subjects toward complex-type N-linked glycosylation in patients. These N-glycosylation shifts were observed on exosomal N-glycoproteins but not on the Tamm-Horsfall glycoprotein, which showed predominant high-mannose-type glycosylation with M6.

• 出版日期2012-2