Figure 6.

Tap oligomers interact with nucleoporins by solid-phase binding assay. (A) Immobilized GST (□) or GST-p62 (■) were incubated with ³⁵S-FLAG-Tap WT or 1-569 deletion mutant. Increasing amounts of recombinant GST-Tap WT or 1-569 were also added to the binding reactions, as indicated. The level of bound radioactive protein was measured by liquid scintillation counting. Binding of ³⁵S-FLAG-Tap WT alone when bound to GST-p62 was arbitrarily assigned a value of 100% and used to normalize the binding values obtained for all other conditions. Each data point represents the mean of three replicates (±SD). (B) Binding of ³⁵S-FLAG-Tap 188-619 and ³⁵S-FLAG-Tap 260-619 to immobilized GST (□) and GST-p62 (■) were compared with that of ³⁵S-FLAG-Tap WT in the presence or absence of recombinant Nxt1. The level of bound radioactive protein was measured by liquid scintillation counting. Each data point represents the mean of three replicates (±SD). (C) Binding of ³⁵S-FLAG-Tap mutant and WT to GST (□) or GST-Nup98-GLFG (■) in the presence or presence of Nxt1 as described in B.