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Article: Proteomic identification and functional characterization of a novel ARF6 GTPase-activating protein, ACAP4

TitleProteomic identification and functional characterization of a novel ARF6 GTPase-activating protein, ACAP4
Authors
Issue Date2006
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc.. The Journal's web site is located at http://www.mcponline.org/
Citation
Molecular And Cellular Proteomics, 2006, v. 5 n. 8, p. 1437-1449 How to Cite?
AbstractARF6 GTPase is a conserved regulator of membrane trafficking and actin-based cytoskeleton dynamics at the leading edge of migrating cells. A key determinant of ARF6 function is the lifetime of the GTP-bound active state, which is orchestrated by GTPase-activating protein (GAP) and GTP-GDP exchanging factor. However, very little is known about the molecular mechanisms underlying ARF6-mediated cell migration. To systematically analyze proteins that regulate ARF6 activity during cell migration, we performed a proteomic analysis of proteins selectively bound to active ARF6 using mass spectrometry and identified a novel ARF6-specific GAP, ACAP4. ACAP4 encodes 903 amino acids and contains two coiled coils, one pleckstrin homology domain, one GAP motif, and two ankyrin repeats. Our biochemical characterization demonstrated that ACAP4 has a phosphatidylinositol 4,5-bisphosphate-dependent GAP activity specific for ARF6. The co-localization of ACAP4 with ARF6 occurred in ruffling membranes formed upon AIF4 and epidermal growth factor stimulation. ACAP4 overexpression limited the recruitment of ARF6 to the membrane ruffles in the absence of epidermal growth factor stimulation. Expression of GTP hydrolysis-resistant ARF6Q67L resulted in accumulations of ACAP4 and ARF6 in the cytoplasmic membrane, suggesting that GTP hydrolysis is required for the ARF6-dependent membrane remodeling. Significantly the depletion of ACAP4 by small interfering RNA or inhibition of ARF6 GTP hydrolysis by overexpressing GAP-deficient ACAP4 suppressed ARF6-dependent cell migration in wound healing, demonstrating the importance of ACAP4 in cell migration. Thus, our study sheds new light on the biological function of ARF6-mediated cell migration. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.
Persistent Identifierhttp://hdl.handle.net/10722/136779
ISSN
2015 Impact Factor: 5.912
2015 SCImago Journal Rankings: 3.213
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorFang, Zen_HK
dc.contributor.authorMiao, Yen_HK
dc.contributor.authorDing, Xen_HK
dc.contributor.authorDeng, Hen_HK
dc.contributor.authorLiu, Sen_HK
dc.contributor.authorWang, Fen_HK
dc.contributor.authorZhou, Ren_HK
dc.contributor.authorWatson, Cen_HK
dc.contributor.authorFu, Cen_HK
dc.contributor.authorHu, Qen_HK
dc.contributor.authorLillard Jr, JWen_HK
dc.contributor.authorPowell, Men_HK
dc.contributor.authorChen, Yen_HK
dc.contributor.authorForte, JGen_HK
dc.contributor.authorYao, Xen_HK
dc.date.accessioned2011-07-29T02:12:07Z-
dc.date.available2011-07-29T02:12:07Z-
dc.date.issued2006en_HK
dc.identifier.citationMolecular And Cellular Proteomics, 2006, v. 5 n. 8, p. 1437-1449en_HK
dc.identifier.issn1535-9476en_HK
dc.identifier.urihttp://hdl.handle.net/10722/136779-
dc.description.abstractARF6 GTPase is a conserved regulator of membrane trafficking and actin-based cytoskeleton dynamics at the leading edge of migrating cells. A key determinant of ARF6 function is the lifetime of the GTP-bound active state, which is orchestrated by GTPase-activating protein (GAP) and GTP-GDP exchanging factor. However, very little is known about the molecular mechanisms underlying ARF6-mediated cell migration. To systematically analyze proteins that regulate ARF6 activity during cell migration, we performed a proteomic analysis of proteins selectively bound to active ARF6 using mass spectrometry and identified a novel ARF6-specific GAP, ACAP4. ACAP4 encodes 903 amino acids and contains two coiled coils, one pleckstrin homology domain, one GAP motif, and two ankyrin repeats. Our biochemical characterization demonstrated that ACAP4 has a phosphatidylinositol 4,5-bisphosphate-dependent GAP activity specific for ARF6. The co-localization of ACAP4 with ARF6 occurred in ruffling membranes formed upon AIF4 and epidermal growth factor stimulation. ACAP4 overexpression limited the recruitment of ARF6 to the membrane ruffles in the absence of epidermal growth factor stimulation. Expression of GTP hydrolysis-resistant ARF6Q67L resulted in accumulations of ACAP4 and ARF6 in the cytoplasmic membrane, suggesting that GTP hydrolysis is required for the ARF6-dependent membrane remodeling. Significantly the depletion of ACAP4 by small interfering RNA or inhibition of ARF6 GTP hydrolysis by overexpressing GAP-deficient ACAP4 suppressed ARF6-dependent cell migration in wound healing, demonstrating the importance of ACAP4 in cell migration. Thus, our study sheds new light on the biological function of ARF6-mediated cell migration. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.en_HK
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc.. The Journal's web site is located at http://www.mcponline.org/en_HK
dc.relation.ispartofMolecular and Cellular Proteomicsen_HK
dc.subject.meshADP-Ribosylation Factors - metabolismen_HK
dc.subject.meshActins - metabolismen_HK
dc.subject.meshAluminum Compounds - chemistryen_HK
dc.subject.meshAmino Acid Substitutionen_HK
dc.subject.meshCell Membrane - metabolismen_HK
dc.subject.meshCell Movement - physiologyen_HK
dc.subject.meshCytoskeleton - physiologyen_HK
dc.subject.meshEpidermal Growth Factor - metabolismen_HK
dc.subject.meshFluorides - chemistryen_HK
dc.subject.meshGTPase-Activating Proteins - chemistry - genetics - metabolismen_HK
dc.subject.meshHeLa Cellsen_HK
dc.subject.meshHumansen_HK
dc.subject.meshIntracellular Membranes - metabolismen_HK
dc.subject.meshMembrane Microdomains - metabolismen_HK
dc.subject.meshPhosphatidylinositol 4,5-Diphosphate - metabolismen_HK
dc.subject.meshProtein Bindingen_HK
dc.subject.meshProteomicsen_HK
dc.titleProteomic identification and functional characterization of a novel ARF6 GTPase-activating protein, ACAP4en_HK
dc.typeArticleen_HK
dc.identifier.emailFu, C:chuanhai@hku.hken_HK
dc.identifier.authorityFu, C=rp01515en_HK
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1074/mcp.M600050-MCP200en_HK
dc.identifier.pmid16737952-
dc.identifier.scopuseid_2-s2.0-33748333436en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33748333436&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume5en_HK
dc.identifier.issue8en_HK
dc.identifier.spage1437en_HK
dc.identifier.epage1449en_HK
dc.identifier.isiWOS:000239627800007-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridFang, Z=14420814200en_HK
dc.identifier.scopusauthoridMiao, Y=8726267000en_HK
dc.identifier.scopusauthoridDing, X=35740254300en_HK
dc.identifier.scopusauthoridDeng, H=49761260800en_HK
dc.identifier.scopusauthoridLiu, S=49761779000en_HK
dc.identifier.scopusauthoridWang, F=14421700300en_HK
dc.identifier.scopusauthoridZhou, R=8353462900en_HK
dc.identifier.scopusauthoridWatson, C=7402883686en_HK
dc.identifier.scopusauthoridFu, C=8583808400en_HK
dc.identifier.scopusauthoridHu, Q=14421200700en_HK
dc.identifier.scopusauthoridLillard Jr, JW=35388902700en_HK
dc.identifier.scopusauthoridPowell, M=35551308000en_HK
dc.identifier.scopusauthoridChen, Y=49761067400en_HK
dc.identifier.scopusauthoridForte, JG=26425932500en_HK
dc.identifier.scopusauthoridYao, X=7402530401en_HK

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