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Figure 8. The BR in the NH2 terminus is required for the ability of PI(4,5)P2 to activate N-WASP. (A) A 10% SDS–polyacrylamide gel showing purified protein preparations of wild-type GST-N-WASP or a GST-tagged mutant of N-WASP containing an internal deletion of the BR (GST-(ΔB)N-WASP). (B) The maximum rate of polymerization was used to quantitate Arp2/3 activity (1.3 μM actin, 30 nM Arp2/3 complex) in the presence of two different concentrations of GST-N-WASP or GST-(ΔB) N-WASP and the indicated combinations of Cdc42-GTPγS (450 nM) and PI(4,5)P2-containing vesicles. While the data shown in B is at 1 μM lipid, GST-(ΔB)N-WASP was PI(4,5)P2-insensitive at higher lipid concentrations as well (up to 5 μM). Image published in: Rohatgi R et al. (2000) © 2000 The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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