Home LiteratureArticle Details
PMID: 22493426 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

IQGAP proteins reveal an atypical phosphoinositide (aPI) binding domain with a pseudo C2 domain fold.

The Journal of biological chemistry ·Vol. 287 ·No. 27 ·2012-06-29 ·Pages 22483-96

Dixon MJ, Gray A, Schenning M, Agacan M, Tempel W, Tong Y, Nedyalkova L, Park HW, Leslie NR, van Aalten DM, Downes CP, Batty IH

Abstract

Class I phosphoinositide (PI) 3-kinases act through effector proteins whose 3-PI selectivity is mediated by a limited repertoire of structurally defined, lipid recognition domains. We describe here the lipid preferences and crystal structure of a new class of PI binding modules exemplified by select IQGAPs (IQ motif containing GTPase-activating proteins) known to coordinate cellular signaling events and cytoskeletal dynamics. This module is defined by a C-terminal 105-107 amino acid region of which IQGAP1 and -2, but not IQGAP3, binds preferentially to phosphatidylinositol 3,4,5-trisphosphate (PtdInsP(3)). The binding affinity for PtdInsP(3), together with other, secondary target-recognition characteristics, are comparable with those of the pleckstrin homology domain of cytohesin-3 (general receptor for phosphoinositides 1), an established PtdInsP(3) effector protein. Importantly, the IQGAP1 C-terminal domain and the cytohesin-3 pleckstrin homology domain, each tagged with enhanced green fluorescent protein, were both re-localized from the cytosol to the cell periphery following the activation of PI 3-kinase in Swiss 3T3 fibroblasts, consistent with their common, selective recognition of endogenous 3-PI(s). The crystal structure of the C-terminal IQGAP2 PI binding module reveals unexpected topological similarity to an integral fold of C2 domains, including a putative basic binding pocket. We propose that this module integrates select IQGAP proteins with PI 3-kinase signaling and constitutes a novel, atypical phosphoinositide binding domain that may represent the first of a larger group, each perhaps structurally unique but collectively dissimilar from the known PI recognition modules.

MeSH Terms
3T3 Cells Amino Acid Sequence Animals Binding Sites/physiology Crystallography GTPase-Activating Proteins/chemistry,genetics,metabolism Mice Molecular Sequence Data Phosphatidylinositol 3-Kinases/metabolism Phosphatidylinositols/metabolism Protein Binding/physiology Protein Structure, Secondary Protein Structure, Tertiary Signal Transduction/physiology ras GTPase-Activating Proteins/chemistry,genetics,metabolism
Chemicals
GTPase-Activating Proteins IQ motif containing GTPase activating protein 1 IQGAP3 protein, mouse Iqgap2 protein, mouse Phosphatidylinositols ras GTPase-Activating Proteins Phosphatidylinositol 3-Kinases
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Dixon Miles J
Division of Cell Signalling and Immunology, College of Life Sciences, University of Dundee, Dow St., Dundee DD1 5EH, Scotland, United Kingdom.
Gray Alexander
Schenning Martijn
Agacan Mark
Tempel Wolfram
Tong Yufeng
Nedyalkova Lyudmila
Park Hee-Won
Leslie Nicholas R
van Aalten Daan M F
Downes C Peter
Batty Ian H
References (50)
50 references, click to expand
  1. Moving towards a better understanding of chemotaxis.
    Curr Biol. 2008 Jun 3;18(11):R485-94 PMID: 18522824
  2. Back in the water: the return of the inositol phosphates.
    Nat Rev Mol Cell Biol. 2001 May;2(5):327-38 PMID: 11331907
  3. Conditional peripheral membrane proteins: facing up to limited specificity.
    Structure. 2012 Jan 11;20(1):15-27 PMID: 22193136
  4. FYVE-finger proteins--effectors of an inositol lipid.
    J Cell Sci. 1999 Dec;112 ( Pt 23):4175-83 PMID: 10564636
  5. Phosphoinositides in cell regulation and membrane dynamics.
    Nature. 2006 Oct 12;443(7112):651-7 PMID: 17035995
  6. Targeting phosphoinositide 3-kinase: moving towards therapy.
    Biochim Biophys Acta. 2008 Jan;1784(1):159-85 PMID: 17997386
  7. A screen for novel phosphoinositide 3-kinase effector proteins.
    Mol Cell Proteomics. 2011 Apr;10(4):M110.003178 PMID: 21263009
  8. Nonradioactive methods for the assay of phosphoinositide 3-kinases and phosphoinositide phosphatases and selective detection of signaling lipids in cell and tissue extracts.
    Anal Biochem. 2003 Feb 15;313(2):234-45 PMID: 12605860
  9. The inhibition of phosphoinositide synthesis and muscarinic-receptor-mediated phospholipase C activity by Li+ as secondary, selective, consequences of inositol depletion in 1321N1 cells.
    Biochem J. 1994 Feb 1;297 ( Pt 3):529-37 PMID: 8110190
  10. Secondary-structure matching (SSM), a new tool for fast protein structure alignment in three dimensions.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2256-68 PMID: 15572779
  11. PTEN: The down side of PI 3-kinase signalling.
    Cell Signal. 2002 Apr;14(4):285-95 PMID: 11858936
  12. Rapid formation of inositol 1,3,4,5-tetrakisphosphate following muscarinic receptor stimulation of rat cerebral cortical slices.
    Biochem J. 1985 Nov 15;232(1):211-5 PMID: 4084229
  13. Structural basis for discrimination of 3-phosphoinositides by pleckstrin homology domains.
    Mol Cell. 2000 Aug;6(2):373-84 PMID: 10983984
  14. Structure of the first C2 domain of synaptotagmin I: a novel Ca2+/phospholipid-binding fold.
    Cell. 1995 Mar 24;80(6):929-38 PMID: 7697723
  15. IQGAPs in cancer: a family of scaffold proteins underlying tumorigenesis.
    FEBS Lett. 2009 Jun 18;583(12):1817-24 PMID: 19433088
  16. Translation of the phosphoinositide code by PI effectors.
    Nat Chem Biol. 2010 Jul;6(7):507-13 PMID: 20559318
  17. Membrane recognition by phospholipid-binding domains.
    Nat Rev Mol Cell Biol. 2008 Feb;9(2):99-111 PMID: 18216767
  18. Membrane recognition and targeting by lipid-binding domains.
    Sci STKE. 2003 Dec 16;2003(213):re16 PMID: 14679290
  19. Signalling through Class I PI3Ks in mammalian cells.
    Biochem Soc Trans. 2006 Nov;34(Pt 5):647-62 PMID: 17052169
  20. Ca(2+) bridges the C2 membrane-binding domain of protein kinase Calpha directly to phosphatidylserine.
    EMBO J. 1999 Nov 15;18(22):6329-38 PMID: 10562545
  21. Structural and mechanistic insights into the association of PKCalpha-C2 domain to PtdIns(4,5)P2.
    Proc Natl Acad Sci U S A. 2009 Apr 21;106(16):6603-7 PMID: 19346474
  22. The pleckstrin homology domains of protein kinase B and GRP1 (general receptor for phosphoinositides-1) are sensitive and selective probes for the cellular detection of phosphatidylinositol 3,4-bisphosphate and/or phosphatidylinositol 3,4,5-trisphosphate in vivo.
    Biochem J. 1999 Dec 15;344 Pt 3:929-36 PMID: 10585883
  23. Refinement of macromolecular structures by the maximum-likelihood method.
    Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55 PMID: 15299926
  24. Phosphoinositide recognition domains.
    Traffic. 2003 Apr;4(4):201-13 PMID: 12694559
  25. IQGAP1: a key regulator of adhesion and migration.
    J Cell Sci. 2005 May 15;118(Pt 10):2085-92 PMID: 15890984
  26. Distinct phosphatidylinositol 3-kinase lipid products accumulate upon oxidative and osmotic stress and lead to different cellular responses.
    J Biol Chem. 1999 Dec 10;274(50):35963-8 PMID: 10585485
  27. Agonist-stimulated synthesis of phosphatidylinositol(3,4,5)-trisphosphate: a new intracellular signalling system?
    Biochim Biophys Acta. 1993 Oct 7;1179(1):27-75 PMID: 8399352
  28. Processing of X-ray diffraction data collected in oscillation mode.
    Methods Enzymol. 1997;276:307-26 PMID: 27754618
  29. The phosphoinositide 3-kinase pathway.
    Science. 2002 May 31;296(5573):1655-7 PMID: 12040186
  30. Metabolic switching of PI3K-dependent lipid signals.
    Biochem Soc Trans. 2007 Apr;35(Pt 2):188-92 PMID: 17371235
  31. A new cytoskeletal connection for APC: linked to actin through IQGAP.
    Dev Cell. 2004 Dec;7(6):778-80 PMID: 15572120
  32. Determining selectivity of phosphoinositide-binding domains.
    Methods. 2006 Jun;39(2):122-33 PMID: 16829131
  33. Signaling by phosphoinositide-3,4,5-trisphosphate through proteins containing pleckstrin and Sec7 homology domains.
    Science. 1997 Mar 28;275(5308):1927-30 PMID: 9072969
  34. The role of phosphatases in inositol signaling reactions.
    J Biol Chem. 1999 Apr 16;274(16):10669-72 PMID: 10196133
  35. IQGAP1 and calmodulin modulate E-cadherin function.
    J Biol Chem. 1999 Dec 31;274(53):37885-92 PMID: 10608854
  36. Regulation of phosphoinositide 3-kinase expression in health and disease.
    Trends Biochem Sci. 2009 Mar;34(3):115-27 PMID: 19299143
  37. IQGAP1 in cellular signaling: bridging the GAP.
    Trends Cell Biol. 2006 May;16(5):242-9 PMID: 16595175
  38. Features and development of Coot.
    Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501 PMID: 20383002
  39. IQGAP1 and its binding proteins control diverse biological functions.
    Cell Signal. 2012 Apr;24(4):826-34 PMID: 22182509
  40. Synthesis and function of 3-phosphorylated inositol lipids.
    Annu Rev Biochem. 2001;70:535-602 PMID: 11395417
  41. Dali: a network tool for protein structure comparison.
    Trends Biochem Sci. 1995 Nov;20(11):478-80 PMID: 8578593
  42. Phosphoinositide 3-kinases in cell migration.
    Biol Cell. 2009 Jan;101(1):13-29 PMID: 19055486
  43. The pleckstrin homology domain of phospholipase C-delta 1 binds with high affinity to phosphatidylinositol 4,5-bisphosphate in bilayer membranes.
    Biochemistry. 1995 Dec 12;34(49):16228-34 PMID: 8519781
  44. The C2 domains of classical/conventional PKCs are specific PtdIns(4,5)P(2)-sensing domains.
    Biochem Soc Trans. 2007 Nov;35(Pt 5):1046-8 PMID: 17956275
  45. Get to grips: steering local actin dynamics with IQGAPs.
    EMBO Rep. 2007 Nov;8(11):1019-23 PMID: 17972901
  46. Dia1 and IQGAP1 interact in cell migration and phagocytic cup formation.
    J Cell Biol. 2007 Jul 16;178(2):193-200 PMID: 17620407
  47. The control of phosphatidylinositol 3,4-bisphosphate concentrations by activation of the Src homology 2 domain containing inositol polyphosphate 5-phosphatase 2, SHIP2.
    Biochem J. 2007 Oct 15;407(2):255-66 PMID: 17672824
  48. In situ proteolysis for protein crystallization and structure determination.
    Nat Methods. 2007 Dec;4(12):1019-21 PMID: 17982461
  49. Automated protein model building combined with iterative structure refinement.
    Nat Struct Biol. 1999 May;6(5):458-63 PMID: 10331874
  50. A new phosphatidylinositol 4,5-bisphosphate-binding site located in the C2 domain of protein kinase Calpha.
    J Biol Chem. 2003 Feb 14;278(7):4972-80 PMID: 12426311
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
Published
2012-06-29
Epub
2012-00-05
Pages
22483-96
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3391087
Subset
IM
Grants
NIGMS NIH HHS · Y1-GM-1104 · United States
Medical Research Council · G9403619 · United Kingdom
Medical Research Council · G0900138 · United Kingdom
CIHR · Canada
Wellcome Trust · 087590 · United Kingdom
Wellcome Trust · 083481 · United Kingdom
NCI NIH HHS · Y1-CO-1020 · United States
Wellcome Trust · United Kingdom
Medical Research Council · G0801865 · United Kingdom
Wellcome Trust · WT087590MA · United Kingdom
Databases
PDB
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]