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. 2000 Sep 18;150(6):1299–1310. doi: 10.1083/jcb.150.6.1299

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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The NH₂-terminal domain of N-WASP does not inhibit binding of G-actin to the VCA segment. GST pull-down assays were used to measure the binding of actin (0.05 μM) to GST-CA, GST-N-WASP, GST-VCA (0.5 μM each), or to GST-VCA (0.5 μM) in the presence of HT-WG or HT-WGP (2.5 μM each). The amount of actin bound was compared using an α-actin immunoblot, loading 100% of the input and 40% of the pulled down material. The amounts of GST-N-WASP, GST-CA, GST-VCA, HT-WG, and HT-WGP pulled down were compared by immunoblotting with α-GST or α-pentaHis.