FIGURE 6.

Ligand‐binding affinity of soluble MET (sMET) with and without N‐glycans. A, Purified recombinant human sMET produced in Flp‐In CHO cells was treated with PNGase F under denaturing conditions or native conditions as described in “Materials and Methods.” The samples were subjected to SDS‐PAGE followed by Coomassie Brilliant Blue R‐250 staining. Left panel, nonreducing PAGE. Right panel, reducing PAGE. B, Binding kinetics of hepatocyte growth factor (HGF) to sMET were evaluated by surface plasmon resonance analysis. HGF was immobilized on a sensor chip C1 and binding of sMET with (right panel) or without (left panel) PNGase F treatment under native condition to HGF was measured using HBS‐EP buffer as described in “Materials and Methods”