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. Author manuscript; available in PMC: 2017 Nov 15.
Published in final edited form as: Biochemistry. 2015 Jul 24;54(30):4711–4730. doi: 10.1021/acs.biochem.5b00362

Figure 8. Binding of CvGal2 to natural glycoproteins.

Figure 8

A) Binding of rCvGal2 to asialofetuin (20 μg/ml) in the presence of serial dilution of asialofetuin (ASF), PSM, asialo-PSM (APSM), OSM, asialo-OSM (AOSM), BSM, or asialo-BSM (ABSM) were measured in solid-based binding assay. Data show optical density at 450 nm (OD450nm) in triplicates with standard error (SEM). IC50 were calculated (Table 3). (B) The binding of CvGal2 to natural glycoproteins was measured in SPR with immobilized rCvGal2, using PSM (left) or asialo-fetuin (right) as analytes. The sensorgrams were recorded with 2-fold serial dilutions starting at the highest concentration lindicated. SPR sensorgram was shown with inset as the 1:1 binding curve. Graphpad Prism software was used to deduce EC50 values. (C) PSM (50 ng/ml) coated wells were treated with neuraminidase (Ctrl) plus N-acetylgalactosidase (GalNac’ase), PNGase F (PNGF), O-glycosidase (OG’ase), or both. Binding of rCvGal2 to treated wells was measured in a solid-phase binding assay. Relative binding to untreated wells was calculated, and the mean with SEM from triplicates is shown. *: p<0.05; ***: p<0.001 vs. control