Identification of ARAP3, a novel PI3K effector regulating both Arf and Rho GTPases, by selective capture on phosphoinositide affinity matrices Journal Article
| Authors: | Krugmann, S.; Anderson, K. E.; Ridley, S. H.; Risso, N.; McGregor, A.; Coadwell, J.; Davidson, K.; Eguinoa, A.; Ellson, C. D.; Lipp, P.; Manifava, M.; Ktistakis, N.; Painter, G.; Thuring, J. W.; Cooper, M. A.; Lim, Z. Y.; Holmes, A. B.; Dove, S. K.; Michell, R. H.; Grewal, A.; Nazarian, A.; Erdjument-Bromage, H.; Tempst, P.; Stephens, L. R.; Hawkins, P. T. |
| Article Title: | Identification of ARAP3, a novel PI3K effector regulating both Arf and Rho GTPases, by selective capture on phosphoinositide affinity matrices |
| 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. |
| Keywords: | signal transduction; controlled study; nonhuman; protein domain; mass spectrometry; proteins; protein analysis; animal cell; animals; animal tissue; protein binding; in vivo study; in vitro study; cos cells; animalia; cloning, molecular; enzyme regulation; molecular sequence data; carrier proteins; recombinant proteins; 1-phosphatidylinositol 3-kinase; adaptor proteins, signal transducing; binding protein; gtpase-activating proteins; cell shape; protein structure; sequence homology; cytoskeleton; rho gtp-binding proteins; swine; cytosol; protein isolation; rho factor; guanosine triphosphatase; cell extract; leukocytes; phosphatidylinositide; tissue extract; adp-ribosylation factors; article; phosphatidylinositol phosphates |
| Journal Title: | Molecular Cell |
| Volume: | 9 |
| Issue: | 1 |
| ISSN: | 1097-2765 |
| Publisher: | Cell Press |
| Date Published: | 2002-01-01 |
| Start Page: | 95 |
| End Page: | 108 |
| Language: | English |
| DOI: | 10.1016/s1097-2765(02)00434-3 |
| PUBMED: | 11804589 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 14 November 2014 -- Source: Scopus |
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