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. Author manuscript; available in PMC: 2021 Jul 7.
Published in final edited form as: Biochem J. 2020 Sep 18;477(17):3433–3451. doi: 10.1042/BCJ20200546

Figure 2. Lysosomes from Arsk-deficient mice do not convert G2A0.

Figure 2.

G2A0 (12.5 nmol) was pre-labeled with the fluorescent dye 2-aminoacridone (AMAC). Fifty micrograms of a lysosome-enriched fraction (tritosomes) were incubated with G2A0-substrate at 37°C for 24 h and subsequently analyzed by C18-reversed-phase-HPLC combined to nano-ESI-MS. For disaccharide code also see [32]. (A) G2A0 was treated with Arsk-deficient tritosomes (blue) resulting in unreacted (gray shaded) substrate 1 (m/z 672.17) or was treated with wildtype (WT) tritosomes. (B) resulting in the novel peak 3 (m/z 416.18). (C) Simultaneous treatment of G2A0 (672.17) with Arsk-deficient tritosomes and human recombinant (r)ARSK (20 ng) resulted in peak 2 (m/z 592.21) indicating the loss of a sulfate group and peak 3 (m/z 416.18) representing the AMAC-labelled hexosamine after desulfation and disaccharide cleavage. (D) Molecular structures and calculated m/z-ratios of the G2A0 disaccharide (1), its desulfated product (2) and the resulting AMAC-labelled monosaccharide product (3), which is due to tritosome-mediated glycosidase activity.