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. 2000 Oct 30;151(3):627–638. doi: 10.1083/jcb.151.3.627

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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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A conserved arginine in ACAP/ASAP family members is necessary for GAP activity. epitope-tagged ACAP1, [R448Q]ACAP1, ACAP2, [R442Q]ACAP2, ASAP1, and [R497K]ASAP1 were expressed and purified as described in Materials and Methods. The GAP activity of ACAP1 and ACAP2, using Arf6 as a substrate, was compared with the GAP activity of an equal amount of [R448Q]ACAP1 and [R442Q]ACAP2. The GAP activity of ASAP1 and [R497K]ASAP1, present at equivalent concentrations, was determined using Arf1 as a substrate. The data were normalized to the activity of wild-type protein, which was 0.05/min for ACAP1, 0.088/min for ACAP2, and 0.35/min for ASAP1. The error bars represent the range of duplicate determinations from one of three experiments.