Figure 2.

N-linked glycosylation at three sites is required for ADA2 secretion. A) Western blot of ADA2 mutants with disrupted N-glycosylation sites. B) Confocal microscopy of ADA2 and ERp72 staining. Arrow indicates perinuclear foci consistent with Golgi apparatus. C) ELISA of ADA2 mutants. D) Enzymatic activity of wild-type and mutant ADA2 quantified using a spectrophotometric assay. ADA activity is coupled to consumption of the substrate NADH measured at OD340. E) Enzymatic activity of ADA2 mutants in the medium of transfected cells normalized to the amount of secreted protein. Data are representative of 3 independent experiments. *p < 0.05.