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. Author manuscript; available in PMC: 2018 Jul 11.
Published in final edited form as: Biochemistry. 2017 Jun 30;56(27):3507–3517. doi: 10.1021/acs.biochem.7b00268

Figure 6.

Figure 6

S-GlcNAcylation of α-synuclein is enzymatically stable. α-Synuclein(gS87) or α-synuclein(gC87) (25 μM, in PBS at pH 7.4) was incubated in triplicate with OGA (1 μM) at 37 °C for 72 h. S-GlcNAc is offset in the y-direction for the sake of clarity. mAU indicates milliabsorbance units. Hydrolysis of GlcNAcylated α-synuclein was analyzed by HPLC at the 72 h time point. Deglycosylation was quantitated by area percent using high-performance liquid chromatography (HPLC) at 214 nm, and the deglycosylated product was characterized by ESI-MS. Results are the mean ± the standard error of the mean of three separate biological experiments. RP-HPLC conditions were 35–60% buffer B over 60 min; buffer A consisted of 0.1% TFA in H2O, and buffer B consisted of 0.1% TFA and 90% ACN in H2O.