FIGURE 6.

Interactions of recombinant globular domains of versican. Solid-phase binding assays were performed using rVN as a soluble ligand (A–C). A, native versican purified from NHDF (●) or BSA (□) was immobilized. Reduced and alkylated rVN (■) was also used as a negative control (indicated as rVN(r)). B, native versican, fibrillin-1 (N) (rF11, fibrillin-1 amino-terminal half), or fibrillin-1 (C) (rF6, fibrillin-1 carboxyl-terminal half) was used to coat wells. Soluble rVN bound to native versican (●). In contrast, rVN did not bind to fibrillin-1 (▵) or fibrillin-1 (C) (×). C, biotin-conjugated rVN bound to wells coated with rVN (♦) and to rVC (recombinant G3 domain, ▴) but not to BSA (□). Binding of soluble biotin-conjugated rVN to immobilized BSA was used as a negative control. Each point in A–C is the mean ± S.D. obtained from three independent experiments. D and E, BIAcore sensorgrams of rVN interactions. Experiments were performed with a series of titrated analytes (rVN) in solution with the indicated concentrations, flowed over a CM5 sensor chip immobilized with 1200 RU of rVN (D) or rF6 (E). The analytes were injected at the point indicated by ▵, and the dissociation phase began at the point indicated by ▴. RU, resonance unit.