Congenital disorders of glycosylation type Ic (CDG-Ic) patients classically present mis-glycosylation of proteins, a diagnostically exploited defect routinely screened for using isoelectric focusing of serum transferrin. To find new biomarkers for identification of this disorder, we screened for mis-glycosylated proteins as a result of lack of a-1,3-glucosyltransferase using S. cerevisiae as a model system Using two-dimensional gel electrophoresis (2-DE), enriched glycoproteins were separated by isoelectric point and protein mass. The glycoprotein spots were excised, trypsin digested, and identified utilizing matrix-assisted laser desorption/ionization tandem time of flight (MALDI-TOF/TOF) mass spectrometry. Compared to the wildtype laboratory strain, mis-glycosidation was found in 17% of the total amount of cytoplasmic elongation factor 2 protein (Eft2p) expressed in a ?alg6 yeast strain. The ability to detect mis-glycosylated Eft2p demonstrated in this study, suggests that similar adverse protein modifications potentially could be identified in humans with CDG type I disorders because the yeast and human elongation factor 2 protein (EF2p) sequences share 66% identity and 85% homology. Future identification of similar mis-glycosylation patterns of EF2p in CDG-I patients could serve as an alternative biomedical diagnostic test for early identification of CDG type I disorders.

Proteomic identification of elongation factor 2 (Eft2p) as a candidate biomarker to characterize protein mis-glycosylation in <i>Saccharomyces cerevisiae</i>

248th National Meeting (2014)

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Proteomic identification of elongation factor 2 (Eft2p) as a candidate biomarker to characterize protein mis-glycosylation in Saccharomyces cerevisiae