FIGURE 2.

Lhs1p, but not Sil1p, requires the linker domain of Kar2p to bind to the nucleotide binding domain. A and B, ability of Sil1p and Lhs1p to bind to the N terminus of Kar2p (NBD + linker), and the NBD alone was assayed. Immobilized GST, GST-Kar2p, GST-NBDlinker (residues 45–438), and GST-NBD (residues 45–425) were incubated with His-Sil1p and His-Lhs1p for 1 h followed by analysis by SDS-PAGE and Coomassie staining. C, binding of both His-Sil1p and His-Lhs1p to GST-NBDlinker was assayed in the presence of 2 mm ADP or 2 mm ATP, and the amount of His-tagged protein bound was analyzed by Western blotting.