A sensitive green fluorescent protein biomarker of N-glycosylation site occupancy.

ABSTRACT: N-glycosylation mediates many biological functions. Genetic defects in the N-glycosylation pathway cause >35 inherited human disorders called congenital disorders of glycosylation (CDGs). As a result, some N-glycosylation sites are unoccupied. Serum transferrin is a diagnostic marker for these patients, but there are no corresponding cellular markers to assess glycosylation competence. Therefore, we engineered a green fluorescent protein (GFP) construct to measure N-glycosylation site occupancy. We designed an endoplasmic reticulum-retained GFP biomarker whose fluorescence is lost when it is N-glycosylated due to steric hindrance by the glycan. This marker is a highly sensitive indicator of N-glycosylation site occupancy. In CDG cells carrying the GFP construct, a 25% decrease of glycosyla

SUBMITTER: Losfeld ME 

PROVIDER: S-EPMC3448770 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature