Figure 1. Sequence determinants of the interaction between the NH2- and COOH-terminal regions of N-WASP. (A) A summary of the nomenclature and amino acid boundaries of the various domains and fragments of bovine N-WASP described in this study. (B) GST-VCA (or GST alone as a control) immobilized on glutathione-Sepharose beads was tested for its ability to pull down various fragments of N-WASP (shown in A) produced as Myc-tagged (MT), 35S-labeled proteins. (C) Dose-dependent binding of the indicated N-WASP fragments to GST-VCA beads. The slope of the line defined by each set of points is inversely proportional to the dissociation constant. (D) GST fusions of the indicated subfragments of the VCA segment of N-WASP (defined in A) were immobilized on glutathione-Sepharose beads and used to pull down the MT-WG fragment from the NH2 terminus of N-WASP. In B and D, 5% of the input and 33% of the pulled down material was analyzed on a 5–15% SDS–polyacrylamide gel.
Image published in: Rohatgi R et al. (2000)
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