Identification of ARAP3, a novel PI3K effector regulating both Arf and Rho GTPases, by selective capture on phosphoinositide affinity matrices

S Krugmann, KE Anderson, SH Ridley, N Risso, A McGregor, J Coadwell, K Davidson, A Eguinoa, CD Ellson, P Lipp, M Manifava, N Ktistakis, G Painter, JW Thuring, MA Cooper, ZY Lim, A Holmes, Stephen Dove, Robert Michell, A GrewalA Nazarian, H Erdjument-Bromage, P Tempst, LR Stephens, PT Hawkins

Research output: Contribution to journalArticlepeer-review

236 Citations (Scopus)

Abstract

We show that matrices carrying the tethered homologs of natural phosphoinositides can be used to capture and display multiple phosphoinositide binding proteins in cell and tissue extracts. We present the mass spectrometric identification of over 20 proteins isolated by this method, mostly from leukocyte extracts: they include known and novel proteins with established phosphoinositide binding domains and also known proteins with surprising and unusual phosphoinositide binding properties. One of the novel PtdIns(3,4,5)P3 binding proteins, ARAP3, has an unusual domain structure, including five predicted PH domains. We show that it is a specific PtdIns(3,4,5)P3/PtdIns(3,4)P2-stimulated Arf6 GAP both in vitro and in vivo, and both its Arf GAP and Rho GAP domains cooperate in mediating PI3K-dependent rearrangements in the cell cytoskeleton and cell shape.
Original languageEnglish
Pages (from-to)95-108
Number of pages14
JournalMolecular Cell
Volume9
Issue number1
DOIs
Publication statusPublished - 1 Jan 2002

Bibliographical note

Copyright © 2002 Cell Press. All rights reserved.

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