title: Classical Galactosemia: Insight into Molecular Pathomechanisms by Differential Membrane Proteomics of Fibroblasts under Galactose Stress.
abstract: Classical galactosemia, a hereditary metabolic disease caused by the deficiency of galactose-1-phosphate uridyltransferase (GALT; EC 2.7.712), results in an impaired galactose metabolism and serious long-term developmental affection of the CNS and ovaries, potentially related in part to endogenous galactose-induced protein dysglycosylation. In search for galactose-induced changes in membrane raft proteomes of GALT-deficient cells, we performed differential analyses of lipid rafts from patient-derived (Q) and sex- and age-matched control fibroblasts (H) in the presence or absence of the stressor. Label-based proteomics revealed of the total 454 (female) or 678 (male) proteins a proportion of ∼12% in at least one of four relevant ratios as fold-changed. GALT(-) cell-specific effects in the absence of stressor revealed cell-model-dependent affection of biological processes related to protein targeting to the plasma membrane (female) or to cellular migration (male). However, a series of common galactose-induced effects were observed, among them the strongly increased ER-stress marker GRP78 and calreticulin involved in N-glycoprotein quality control. The membrane-anchored N-glycoprotein receptor CD109 was concertedly decreased under galactose-stress together with cadherin-13, GLIPR1, glypican-1, and semaphorin-7A. A series of proteins showed opposite fold-changes in the two cell models, whereas others fluctuated in only one of the two models.
authors.affiliation: Institute of Biochemistry II, Medical Faculty, University of Cologne , Joseph-Stelzmann-Str. 52, 50931 Köln, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Staubach
authors.affiliation: Center for Molecular Medicine Cologne, University of Cologne , Robert-Koch-Str. 21, 50931 Köln, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Müller
authors.affiliation: Institute of Biochemistry II, Medical Faculty, University of Cologne , Joseph-Stelzmann-Str. 52, 50931 Köln, Germany.|Department of Molecular Biology and Genetics, Faculty of Science, Istanbul University , Istanbul 34134, Turkey.
authors.forename: Murat
authors.initials: M
authors.lastname: Pekmez
authors.affiliation: Institute of Biochemistry II, Medical Faculty, University of Cologne , Joseph-Stelzmann-Str. 52, 50931 Köln, Germany.|Center for Molecular Medicine Cologne, University of Cologne , Robert-Koch-Str. 21, 50931 Köln, Germany.
authors.forename: Franz-Georg
authors.identifier: 0000-0003-3619-7039
authors.initials: FG
authors.lastname: Hanisch
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D037282:Calreticulin / Q000235:genetics / Q000378:metabolism; D016022:Case-Control Studies; D002675:Child, Preschool; D000091342:Endoplasmic Reticulum Chaperone BiP; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism; D005690:Galactose / Q000494:pharmacology*; D005693:Galactosemias / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D063990:Gene Ontology; D053673:Glypicans / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007223:Infant; D008297:Male; D021962:Membrane Microdomains / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D008565:Membrane Proteins; D058977:Molecular Sequence Annotation; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D039961:Semaphorins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013312:Stress, Physiological; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase
chemical_list: D015703:Antigens, CD; C576483:CALR protein, human; C120523:CD109 protein, human; D015820:Cadherins; D037282:Calreticulin; D000091342:Endoplasmic Reticulum Chaperone BiP; C094538:GLIPR1 protein, human; D058851:GPI-Linked Proteins; D053673:Glypicans; C099972:H-cadherin; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; C118753:SEMA7A protein, human; D039961:Semaphorins; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase; D005690:Galactose
keywords: GALT deficiency; classical galactosemia; differential proteomics; iTRAQ; lipid rafts; membrane glycoproteins