title: A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2.
abstract: Using a series of immunoprecipitation (IP)-tandem mass spectrometry (LC-MS/MS) experiments and reciprocal BLAST, we conducted a fly-human cross-species comparison of the phosphoinositide-3-kinase (PI3K) interactome in a drosophila S2R+ cell line and several NSCLC and human multiple myeloma cell lines to identify conserved interacting proteins to PI3K, a critical signaling regulator of the AKT pathway. Using H929 human cancer cells and drosophila S2R+ cells, our data revealed an unexpected direct binding of Corkscrew, the drosophila ortholog of the non-receptor protein tyrosine phosphatase type II (SHP2) to the Pi3k21B (p60) regulatory subunit of PI3K (p50/p85 human ortholog) but no association with Pi3k92e, the human ortholog of the p110 catalytic subunit. The p85-SHP2 association was validated in human cell lines, and formed a ternary regulatory complex with GRB2-associated-binding protein 2 (GAB2). Validation experiments with knockdown of GAB2 and Far-Western blots proved the direct interaction of SHP2 with p85, independent of adaptor proteins and transfected FLAG-p85 provided evidence that SHP2 binding on p85 occurred on the SH2 domains. A disruption of the SHP2-p85 complex took place after insulin/IGF1 stimulation or imatinib treatment, suggesting that the direct SHP2-p85 interaction was both independent of AKT activation and positively regulates the ERK signaling pathway.
authors.affiliation: Beth Israel Deaconess Medical Center, Division of Signal Transduction, Boston, MA USA.|Harvard Medical School, Department of Medicine, Boston, MA USA.
authors.forename: Susanne B
authors.initials: SB
authors.lastname: Breitkopf
authors.affiliation: Beth Israel Deaconess Medical Center, Division of Signal Transduction, Boston, MA USA.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Yang
authors.affiliation: Beth Israel Deaconess Medical Center, Division of Signal Transduction, Boston, MA USA.|Centers for Therapeutic Innovation, Pfizer, Boston, Massachusetts, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Begley
authors.affiliation: Howard Hughes Medical Institute, Harvard Medical School, Department of Genetics, Boston, MA USA.
authors.forename: Meghana
authors.initials: M
authors.lastname: Kulkarni
authors.affiliation: Novartis, Cambridge, MA USA.
authors.forename: Yu-Hsin
authors.initials: YH
authors.lastname: Chiu
authors.affiliation: Massachusetts General Hospital, Center for Thoracic Cancers, Charlestown, MA USA.
authors.forename: Alexa B
authors.initials: AB
authors.lastname: Turke
authors.affiliation: Beth Israel Deaconess Medical Center, Division of Cardiology, Boston, MA USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Lauriol
authors.affiliation: Beth Israel Deaconess Medical Center, Division of Signal Transduction, Boston, MA USA.
authors.forename: Min
authors.initials: M
authors.lastname: Yuan
authors.affiliation: Massachusetts General Hospital, Center for Thoracic Cancers, Charlestown, MA USA.
authors.forename: Jie
authors.initials: J
authors.lastname: Qi
authors.affiliation: Massachusetts General Hospital, Center for Thoracic Cancers, Charlestown, MA USA.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Engelman
authors.affiliation: Brandeis University, Department of Computer Science, Waltham, MA USA.
authors.forename: Pengyu
authors.initials: P
authors.lastname: Hong
authors.affiliation: Beth Israel Deaconess Medical Center, Division of Cardiology, Boston, MA USA.|Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA USA.
authors.forename: Maria I
authors.initials: MI
authors.lastname: Kontaridis
authors.affiliation: Cancer Center, Weill Cornell Medical College, New York, NY USA.
authors.forename: Lewis C
authors.initials: LC
authors.lastname: Cantley
authors.affiliation: Howard Hughes Medical Institute, Harvard Medical School, Department of Genetics, Boston, MA USA.
authors.forename: Norbert
authors.initials: N
authors.lastname: Perrimon
authors.affiliation: Beth Israel Deaconess Medical Center, Division of Signal Transduction, Boston, MA USA.|Harvard Medical School, Department of Medicine, Boston, MA USA.
authors.forename: John M
authors.initials: JM
authors.lastname: Asara
mesh_terms: D000818:Animals; D002460:Cell Line; D004175:Diptera / Q000378:metabolism*; D004330:Drosophila / Q000378:metabolism*; D029721:Drosophila Proteins / Q000378:metabolism; D006801:Humans; D019476:Insect Proteins / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011485:Protein Binding; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11 / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D029721:Drosophila Proteins; D019476:Insect Proteins; D019869:Phosphatidylinositol 3-Kinases; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11
references.citation: Circ Res. 2003 Aug 8;93(3):221-9
references.pmid: 12855672
references.citation: Science. 2002 May 31;296(5573):1655-7
references.pmid: 12040186
references.citation: Biochem Biophys Res Commun. 2004 Oct 8;323(1):345-54
references.pmid: 15351743
references.citation: Nat Genet. 2001 Dec;29(4):465-8
references.pmid: 11704759
references.citation: J Immunol. 2004 Oct 15;173(8):4953-9
references.pmid: 15470037
references.citation: Acta Diabetol. 1992;28(3-4):215-20
references.pmid: 1576358
references.citation: EMBO J. 1992 Sep;11(9):3469-79
references.pmid: 1380456
references.citation: Mol Cell Biol. 1994 Apr;14(4):2777-85
references.pmid: 7511210
references.citation: J Biol Chem. 1994 Apr 29;269(17):12503-7
references.pmid: 8175658
references.citation: J Biol Chem. 1994 Sep 23;269(38):23764-8
references.pmid: 7522233
references.citation: Methods Enzymol. 1995;254:523-35
references.pmid: 8531712
references.citation: J Biol Chem. 1997 Nov 14;272(46):29281-9
references.pmid: 9361008
references.citation: Mol Cell. 1998 Dec;2(6):729-40
references.pmid: 9885561
references.citation: Biochem Biophys Res Commun. 1999 Jan 19;254(2):440-5
references.pmid: 9918857
references.citation: Cell. 1999 Jun 25;97(7):865-75
references.pmid: 10399915
references.citation: Bioinformatics. 2003 Sep 1;19(13):1710-1
references.pmid: 15593400
references.citation: Dev Dyn. 2005 Mar;232(3):827-34
references.pmid: 15704125
references.citation: Methods Mol Biol. 2005;309:1-9
references.pmid: 15990393
references.citation: J Biol Chem. 2005 Jul 29;280(30):27850-5
references.pmid: 15932879
references.citation: J Cell Biol. 2005 Aug 1;170(3):455-64
references.pmid: 16043515
references.citation: EMBO J. 2006 Oct 18;25(20):4740-51
references.pmid: 17024187
references.citation: BMC Bioinformatics. 2006;7:457
references.pmid: 17044912
references.citation: J Biol Chem. 2007 Mar 16;282(11):7758-69
references.pmid: 17178724
references.citation: Science. 2007 Jul 13;317(5835):239-42
references.pmid: 17626883
references.citation: Nat Protoc. 2007;2(12):3278-84
references.pmid: 18079728
references.citation: Science. 2007 Dec 14;318(5857):1744-8
references.pmid: 18079394
references.citation: Bioinformatics. 2008 Feb 1;24(3):319-24
references.pmid: 18042555
references.citation: PLoS One. 2008;3(2):e1562
references.pmid: 18253506
references.citation: Proteins. 2008 May 15;71(3):1113-22
references.pmid: 18004756
references.citation: Oncogene. 2008 Sep 18;27(41):5486-96
references.pmid: 18794883
references.citation: Cell Cycle. 2008 Oct;7(19):2991-6
references.pmid: 18818526
references.citation: Curr Protoc Mol Biol. 2012 Apr;Chapter 18:Unit18.19.1-27
references.pmid: 22470061
references.citation: J Biol Chem. 2002 May 31;277(22):19382-8
references.pmid: 11896055
references.citation: Sci Signal. 2011;4(196):rs10
references.pmid: 22028469
references.citation: J Med Genet. 2002 Aug;39(8):571-4
references.pmid: 12161596
references.citation: Endocrine. 2002 Jun;18(1):1-12
references.pmid: 12166618
references.citation: Proc Natl Acad Sci U S A. 2009 May 5;106(18):7531-6
references.pmid: 19380737
references.citation: Mol Cell Biol. 2009 Oct;29(19):5377-88
references.pmid: 19635806
references.citation: J Biomol Tech. 2009 Dec;20(5):272-7
references.pmid: 19949701
references.citation: Sci Signal. 2009 Dec 8;2(100):re9
references.pmid: 19996458
references.citation: Proc Natl Acad Sci U S A. 2010 Mar 23;107(12):5471-6
references.pmid: 20212113
references.citation: Curr Top Microbiol Immunol. 2010;346:31-56
references.pmid: 20517722
references.citation: Proc Natl Acad Sci U S A. 2012 Oct 2;109(40):16190-5
references.pmid: 22988110
references.citation: PLoS One. 2012;7(10):e46683
references.pmid: 23071613
references.citation: PLoS Genet. 2012;8(12):e1003161
references.pmid: 23300466
references.citation: Sci Signal. 2013 May 21;6(276):ra38
references.pmid: 23695164
references.citation: Mol Cell. 2014 Apr 10;54(1):193-202
references.pmid: 24657164
references.citation: Exp Cell Res. 1999 Nov 25;253(1):239-54
references.pmid: 10579926
references.citation: J Biol Chem. 2001 Jan 26;276(4):2494-502
references.pmid: 11054408
references.citation: Curr Cancer Drug Targets. 2008 Dec;8(8):733-40
references.pmid: 19075596
references.citation: Analyst. 2009 Apr;134(4):755-62
references.pmid: 19305927
references.citation: Mol Cell Biol. 2002 Jun;22(12):4062-72
references.pmid: 12024020
references.citation: Mol Cell. 2011 Mar 4;41(5):567-78
references.pmid: 21362552
references.citation: Cancer Res. 2011 Sep 15;71(18):5965-75
references.pmid: 21775521
references.citation: J Biol Chem. 2003 Nov 28;278(48):48453-66
references.pmid: 14504291