research article signaling network map of endothelial tek … · 2019. 7. 31. · tie interacts...

Research ArticleSignaling Network Map of Endothelial TEK Tyrosine Kinase

Aafaque Ahmad Khan12 Varot K Sandhya1 Priyata Singh3 Deepak Parthasarathy3

Awinav Kumar3 Jayshree Advani14 Rudrappa Gattu12 Dhanya V Ranjit1

Rama Vaidyanathan3 Premendu Prakash Mathur2 T S Keshava Prasad1

F Mac Gabhann5 Akhilesh Pandey16789 Rajesh Raju1 and Harsha Gowda1

1 Institute of Bioinformatics International Tech Park Whitefield Bangalore 560066 India2 School of Biotechnology KIIT University Bhubaneswar 751024 India3 Dr MGR Educational and Research Institute Maduravoyal Chennai 600095 India4Manipal University Madhav Nagar Manipal 576104 India5 Department of Biomedical Engineering and Institute for Computational Medicine Johns Hopkins University BaltimoreMD 21218 USA

6McKusick-Nathans Institute of Genetic Medicine Johns Hopkins University School of Medicine Baltimore MD 21205 USA7Department of Biological Chemistry Johns Hopkins University School of Medicine Baltimore MD 21205 USA8Department of Pathology Johns Hopkins University School of Medicine Baltimore MD 21205 USA9Department of Oncology Johns Hopkins University School of Medicine Baltimore MD 21205 USA

Correspondence should be addressed to Rajesh Raju rajeshibioinformaticsorg and Harsha Gowda harshaibioinformaticsorg

Received 22 April 2014 Accepted 15 September 2014 Published 13 October 2014

Academic Editor Shoukat Dedhar

Copyright copy 2014 Aafaque Ahmad Khan et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

TEK tyrosine kinase is primarily expressed on endothelial cells and is most commonly referred to as TIE2 TIE2 is a receptortyrosine kinase modulated by its ligands angiopoietins to regulate the development and remodeling of vascular system It is alsoone of the critical pathways associatedwith tumor angiogenesis and familial venousmalformations Apart from the vascular systemTIE2 signaling is also associated with postnatal hematopoiesis Despite the involvement of TIE2-angiopoietin system in severaldiseases the downstream molecular events of TIE2-angiopoietin signaling are not reported in any pathway repository Thereforecarrying out a detailed review of published literature we have documentedmolecular signaling eventsmediated byTIE2 in responseto angiopoietins and developed a network map of TIE2 signaling The pathway information is freely available to the scientificcommunity through NetPath a manually curated resource of signaling pathways We hope that this pathway resource will providean in-depth view of TIE2-angiopoietin signaling andwill lead to identification of potential therapeutic targets for TIE2-angiopoietinassociated disorders

1 Introduction

Angiopoietin-TIE2 is one of the major signaling systems thatregulates development and remodeling of vascular system[1 2] TIE2 is a member of the TIE receptor tyrosine kinasefamily that is preferentially expressed in endothelial cells[3] Among the angiopoietins (angiopoietin-1 angiopoietin-2 and angiopoietin-4 in humans) angiopoietin-1 (ANGPT1)is known as a constitutive agonist of TIE2 ANGPT1TIE2signaling promotes endothelial cell survival endothelium

integrity and anti-inflammatoryantiapoptotic responsessupporting reduced vascular permeability [4 5] ANGPT2is generally considered as antagonist as it competes withANGPT1 for binding to TIE2 reduces vessel stability andenhances vascular remodeling [6] However under spe-cific experimental conditions ANGPT2 has been shown topromote endothelial-cell survival sprouting and migrationin a temporal and concentration-dependent manner [7ndash9]Therefore angiopoietin-2 (ANGPT2) is currently consideredas a context dependant agonist or antagonist of TIE2 [6 10]

Hindawi Publishing CorporationJournal of Signal TransductionVolume 2014 Article ID 173026 6 pageshttpdxdoiorg1011552014173026

2 Journal of Signal Transduction

Angiopoietin-4 (ANGPT4) is also known to be an agonist ofTIE2while angiopoietin-3 (ANGPT3) themouse ortholog ofangiopoietin-4 is reported to be antagonistic toTIE2 [11]Theother member of the TIE family is the orphan receptor TIE1It heterodimerizes with TIE2 and modulates TIE2 signalinginduced by ANGPT1 and ANGPT2 [12] ANGPT1 binding toTIE2 induces dissociation of the TIE1-TIE2 complex [12]Thissuggests that TIE2 signaling is regulated by the molecularbalance between ANGPT1 and ANGPT2 [6 13] and TIE1and TIE2 with another one being the ectodomain cleavageof TIE receptors [14] The activation of TIE2 is achievedby the assembly with tetrameric or higher order multimericangiopoietins clearly differentiating TIE2 from other tyro-sine kinase receptors [15] ANGPT1 induces the translocationof TIE2 to cell-cell junctions and transassociation in theform of homomeric complexes to activate the downstreamsignaling of TIE2 [16]

Binding of ANGPT1 to TIE2 leads to receptor dimer-ization and subsequent activation followed by autophospho-rylation at specific tyrosine residues [15 17] These phos-phorylated sites provide binding platform to a number ofeffector molecules to initiate downstream signaling cascadewhich ultimately controls various cellular responses includ-ing morphogenesis proliferation extracellular matrix inter-action permeability survival and differentiation [18ndash23]TIE2 interacts with p85 subunit of phosphatidylinositol-3-kinase (PI3K) via Tyr-1101 and activates PI3K-AKT pathwaywhich inhibits Smac release frommitochondria and increasesthe expression of survivin leading to survival and chemo-taxis of endothelial cells [18 24 25] AKT activation alsoinhibits forkhead transcription factor FKHR (FOXO1) whichprotects endothelial cells from apoptosis [26] ANGPT1 alsoinduces the PI3KAKT mediated activation of eNOS andNO release in endothelial cells [27 28] In endothelial cellsboth ANGPT1 and ANGPT2 also induce TIE2-dependenttranslocation of P-selectin through a PLCG1Ca2+ signalingpathway [29]

SH2 domain containing proteins such as growth factorreceptor-bound protein 2 (GRB2) growth factor receptor-bound protein 7 (GRB7) growth factor receptor-bound pro-tein 14 (GRB14) protein tyrosine phosphatase nonreceptortype 11 (SHP-2) and phosphoinositide-3-kinase (PI3K) isrecruited and transphosphorylated by TIE2 [30] GRB2 andSHC1 recruit SOS1 and lead to the activation of Ras-Raf-mitogen activated protein kinase (MAPK) pathway that reg-ulates platelet activating factor synthesis anti-inflammatoryresponses and endothelial cell migration proliferation per-meability and morphogenesis [5 20ndash22 31 32] ThroughSOS1 or PI3Ks angiopoietinTIE2 system also regulates theactivation of RAC1 RHOA CDC42 and focal adhesionkinase 1 to mediate cytoskeleton reorganization and migra-tion of endothelial and synovial cells [33] Angiopoietin-1 induced activation of RHOA results in sequestration ofSRC by DIAPH1 thereby preventing SRC association withVEGFR2 [34] Recruitment of dynamic complexes com-prising NCK adaptor protein 1 (NCK1) RAS p21 proteinactivator 1 (p120GAP) and P21 protein-activated kinase 1(PAK1) to TIE2 by the DOKs especially DOK2 has beenattributed to increased cell motility [35] TIE2 also interacts

with the inhibitor of nuclear factor kappa B (NF-kB) activityTNFAIP3 interacting protein 2 (ABIN-2) that inhibits NF-kB transcriptional activity and mediates anti-inflammatoryand antiapoptotic action [36 37] TIE2 activation inducesthe phosphorylation of STAT1 STAT3 and STAT5A5B andtheir subsequent translocation into nucleus to induce theexpression of the cell cycle inhibitor cyclin-dependent kinaseinhibitor 1A (p21) [38] ANGPT2 also interacts with integrinslike integrin 120572V1205735 120572V1205733 and 12057251205731 in endothelial cells withless affinity than TIE2 and can induce TIE2-independentsignaling [39] TIE2 also forms a complex with 12057251205733 andFAKANGPT2 induces phosphorylation of FAK at Serine91012057251205733 internalization and dissociation of p130CAS and talinfrom 12057251205733 [40] Recently ANGPT2 has also been shown toinduce the activation of ERKMSK1CREB pathway to impartcell survival and resistance to doxorubicin in HepG2 cells[41]

Besides the defects in vascular system and angiogene-sis [42ndash44] TIE2 signaling has also been associated withrheumatoid arthritis [45] and asthma [46] Considering theimportance of TIE2 signaling here we provide a manuallycurated enhanced network map of angiopoietin(s)-inducedTIE2-mediated signaling events as a reference platform forfurther biomedical investigations

2 Methods

We screened published research articles related to TIE2signaling NetPath criteria described earlier [47 48] werefollowed for the annotation of protein-protein interac-tions (PPIs) enzyme-substrate relationships and posttrans-lational modifications (PTMs) (catalytic events) Activa-tioninhibition status of proteins alterations in protein local-ization and also genes regulated at mRNA level by TIE2signaling were also documented PathBuilder an in-housepathway annotation tool was used for the curation of thesereactions [49] Each curated reaction was internally reviewedby trained biocurators followed by an external review by aPathway Authority an expert in the field (FMG coauthor ofthis paper)

3 Results and Discussion

Our analysis resulted in the cataloging of 140 uniquemolecules that are reported in TIE2 signaling Thesemolecules were part of 43 PPIs and 102 catalytic events23 activationinhibition events and 11 protein transloca-tion events We have also documented 124 and 65 genesthat were reported to be upregulated and downregulatedrespectively by TIE2 signaling in response to angiopoi-etin(s) in human cells The curated data for TIE2 sig-naling pathway is freely available to the scientific com-munity for visualization and download in different com-munity standard data exchange formats through NetPath[httpwwwnetpathorg] a resource of signaling path-ways These formats include Proteomics Standards Initiativefor Molecular Interaction (PSI-MI version 21) Biological

Journal of Signal Transduction 3

PAK1S204T423

STAT5BY699

STAT5AY694

AKT2S474

STAT5BY699

MAPK9T183 Y185

STAT5AY694

STAT3Y705

TEK

PIK3CA

BCAR1 P

PIK3R2P

GRB14 P

TEKY1108TEK Y1102 Y992Y1113

ANGPT1

ANGPT1

ANGPT1ANGPT1

CRK

RASA1

DOK4

Fibronectin

NCK1P

DOK2Y351

Rearrangement ofactin cytoskeleton

PIK3R1 P

GRB2

GRB7P

PTPN11P

ITGA5

PTK2Y861 S910

Y397

ITGB1

MAPK1Y187T185

MAPK3Y204T202MMP2

ILK

AKT1 T308S473GSK3B S9Endothelial

cell migration

RAC1GTP

NOS3S1179

TNIP2

PTPRB

ANGPT2

TEKP

CDKN1A

Endothelial cellcycle arrest

Oligomeric

MTORS2481

HIF1A

CXCL12

Neovasculogenesisand protection

against ischemia inendothelial cells

CBL

PLD2PLD1

RASGTP

RAF1P

MAP2K1S218

MAP2K2S222

NFKBIAP

Dissociation

RELA

RELA

CY

NU

Degradation

PDPK1P

PIP2

PIP3

ROCK1

CDC42GTP

RHOAGTP

DIAPH1

Synovial cell migration

Synovial cell growth

FOXO1S256T24S319

GJA5

Vascular quiescencein human vascular

endothelial cells

FOXO1S319T24

S256

CY

NU

SELP

PLCG1

SELP

PKC

Ca2+

Proinflammatoryresponse

NR4A1

Anti-inflammatoryresponses

PXN P

PLG

FESP

FYN P

Tube formationin brain capillaryendothelial cells

RPS6KB1T389S411

EGR1

SOS1

ARHGAP5 P

MAPK14Y182T180

RPS6KA1S380

ELK1S383

MAP3K3S526

MAPK7 P

Endothelial cellproliferation

Platelet activating factor

synthesis

PLA2G5

Phospholipids

Lyso-PAF

Dissociation

DLL4

CTNNB1

MAPK8Y185 T183

BCL2

Cell survivaland migration

FOXO3S253 S318S321

BIRC5

Antiapoptosis in microvascular

and retinalendothelial cells

MAP2K4S80

STAT3 Y705

STAT1 Y701

SPHK1P

Inhibition ofendothelial cell

permeability

Endothelialprogenitor cell angiogenesis

HSP90AA1

ITGB3ITGAV

Ub

Internalizationubiquitination

and degradation

Dissociation

TIE-2 signaling pathway

SHC1P

SRC

TIE1 P

BMXP

Promotes endothelialbarrier defense in

human MECs

ROS

NOX

NO


endothelial cells

TEK

Angiopoietin-1 bridgedTIE2 transassociationand cell-cell contacts

Extracellularmatrix boundangiopoietin-1

MEF2A

MEF2C

MEF2D

EGR1

Endothelial cellmigration

proliferation andcapillary-like tube

formation

Internalizationand degradation

Dissociation

TLN1

VEGFA

KLF2

CY

NU

EGR1

CY

NU

STAT1Y701

IL8

JUNY182T180

Endothelial cellmigration andproliferation

MAPK14T180Y182

ANGPT4

TEK TEK

ANGPT2 ANGPT4

Dissociation ITGA5ITGB1 ITGB3

ITGAVITGB5ITGAV

ANGPT2 ANGPT2 ANGPT2

Preventsbinding of

SRC to VEGFR2

Dissociation

PLG

TIAM1

PRKCZ

CTNNB1ZAP70P

EGFRP

ITGB2 P

GNA11 P

PGK1 P

PDIA3 P

ALDOA P

GTPBP3 P

HAGHP

MASP2 P

ADSS P

TXNRD3P

TXNRD1P CTPS1 P

GYS1 P

LIMCH1P

ProteinReceptor

Protein

Protein

mRNA

Small molecule

Ligand

Induced activation

Enzyme complex

Inhibition

Autocatalysis

Transport

Positive regulation of gene expression

Leads to through unknown mechanism

Negative regulation of gene expression

Translocation

Ubiquitination

PTM state

Protein-protein interaction

Dephosphorylation

Phosphorylation

NucleusCytoplasmPlasma membrane

CYPM

NU

Molecule

ANGPT2Inhibition of

FOXO1 leads tosuppression of

ANGPT2expression

IKK

Showing direction of line

Figure 1 A detailed map of TIE2 signaling This is a manually drawn (using PathVisio) pictorial representation of the network of reactionsannotated in NetPath The topology of the molecules and their reactions from TIE2 to the transcription factors are derived from theexperimental information obtained by the use of inhibitors activatorsmutants and silencing approaches Each node represents themoleculesand the edges represent the relationships between them as provided in the figure legend


Pathway Exchange (BioPAX level 3) and Systems BiologyMarkup Language (SBML level 21)

For effective visualization we have graphically repre-sented the reactions and various cellular processes thatthose reactions mediate in the context of specific studieson TIE2 signaling (Figure 1) PathVisio an open visual-ization tool was used to manually depict this information[50] The pathway map can also be accessed through Net-Slim (httpwwwnetpathorgnetslimTIE2 pathwayhtml)a resource that provides a smaller version of the pathwayby filtering data based on predefined confidence thresholdcriteria [51] At NetSlim a ldquomap with citationrdquo is also pro-vided in which each reaction is linked to the correspondingliterature through PubMed Users can download these mapsin customizable formats such as GenMAPP and gpml

4 Conclusions

This open-access pathway data enables better analysis ofhigh-throughput experimental data and hypothesis-drivenapproaches to study the dynamics of TIE2 signaling fortherapeutic interventions Information on TIE2 pathwayin NetPath will be periodically updated to reflect novelfindings relevant to TIE2 signaling We intend to pro-vide information pertaining to cross-talks of other lig-andreceptor systems such as VEGF TNF-alpha and inte-grins with TIE2 and vice versa in the subsequent versionsin NetPath We encourage scientific community to help usmaintain this resource up-to-date and error-free throughhttpwwwnetpathorgcomments

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors thank the Department of Biotechnology (DBT)Government of India for research support to the Instituteof Bioinformatics Bangalore Harsha Gowda is a WellcomeTrustDBT India Alliance Early Career Fellow

References

[1] D J Dumont G Gradwohl G-H Fong et al ldquoDominant-negative and targeted null mutations in the endothelial receptortyrosine kinase tek reveal a critical role in vasculogenesis of theembryordquo Genes and Development vol 8 no 16 pp 1897ndash19091994

[2] A L Wong Z A Haroon S Werner M W Dewhirst C SGreenberg and K G Peters ldquoTie2 expression and phospho-rylation in angiogenic and quiescent adult tissuesrdquo CirculationResearch vol 81 no 4 pp 567ndash574 1997

[3] D J Dumont T P Yamaguchi R A Conlon J Rossant andM L Breitman ldquoTek a novel tyrosine kinase gene located onmouse chromosome 4 is expressed in endothelial cells and theirpresumptive precursorsrdquo Oncogene vol 7 no 8 pp 1471ndash14801992

[4] U Fiedler T Krissl S Koidl et al ldquoAngiopoietin-1 andangiopoietin-2 share the same binding domains in the Tie-2 receptor involving the first Ig-like loop and the epidermalgrowth factor-like repeatsrdquoThe Journal of Biological Chemistryvol 278 no 3 pp 1721ndash1727 2003

[5] C Suri J McClain G Thurston et al ldquoIncreased vasculariza-tion in mice overexpressing angiopoietin-1rdquo Science vol 282no 5388 pp 468ndash471 1998

[6] P C Maisonpierre C Suri P F Jones et al ldquoAngiopoietin-2 anatural antagonist for Tie2 that disrupts in vivo angiogenesisrdquoScience vol 277 no 5322 pp 55ndash60 1997

[7] I Kim J-H Kim S-O Moon H J Kwak N-G Kim andG Y Koh ldquoAngiopoietin-2 at high concentration can enhanceenthelial cell survival through the phosphatidylinositol 31015840-kinaseAkt signal transduction pathwayrdquo Oncogene vol 19 no39 pp 4549ndash4552 2000

[8] Y Mochizuki T Nakamura H Kanetake and S KandaldquoAngiopoietin 2 stimulates migration and tube-like structureformation of murine brain capillary endothelial cells throughc-Fes and c-Fynrdquo Journal of Cell Science vol 115 part 1 pp 175ndash183 2002

[9] K Teichert-Kuliszewska P C Maisonpierre N Jones et alldquoBiological action of angiopoietin-2 in a fibrin matrix model ofangiogenesis is associated with activation of Tie2rdquoCardiovascu-lar Research vol 49 no 3 pp 659ndash670 2001

[10] S Davis T H Aldrich P F Jones et al ldquoIsolation ofangiopoietin-1 a ligand for the TIE2 receptor by secretion-trapexpression cloningrdquo Cell vol 87 no 7 pp 1161ndash1169 1996

[11] D M Valenzuela J A Griffiths J Rojas et al ldquoAngiopoietins3 and 4 diverging gene counterparts in mice and humansrdquoProceedings of the National Academy of Sciences of the UnitedStates of America vol 96 no 5 pp 1904ndash1909 1999

[12] T C M Seegar B Eller D Tzvetkova-Robev et al ldquoTie1-Tie2interactions mediate functional differences between angiopoi-etin ligandsrdquoMolecular Cell vol 37 no 5 pp 643ndash655 2010

[13] H T Yuan E V Khankin S A Karumanchi and S M ParikhldquoAngiopoietin 2 is a partial agonistantagonist of Tie2 signalingin the endotheliumrdquoMolecular and Cellular Biology vol 29 no8 pp 2011ndash2022 2009

[14] H Singh T M Hansen N Patel and N P J Brindle ldquoThemolecular balance between receptor tyrosine kinases Tie1 andTie2 is dynamically controlled by VEGF and TNF120572 and regu-lates angiopoietin signallingrdquo PLoS ONE vol 7 no 1 Article IDe29319 2012

[15] S Davis N Papadopoulos T H Aldrich et al ldquoAngiopoietinshave distinct modular domains essential for receptor bindingdimerization and superclusteringrdquo Nature Structural Biologyvol 10 no 1 pp 38ndash44 2003

[16] S Fukuhara K Sako T Minami et al ldquoDifferential function ofTie2 at cell-cell contacts and cell-substratum contacts regulatedby angiopoietin-1rdquo Nature Cell Biology vol 10 no 5 pp 513ndash526 2008

[17] K-T Kim H-H Choi M O Steinmetz et al ldquoOligomerizationand multimerization are critical for angiopoietin-1 to bind andphosphorylate Tie2rdquo Journal of Biological Chemistry vol 280no 20 pp 20126ndash20131 2005

[18] A Brkovic M Pelletier D Girard andM G Sirois ldquoAngiopoi-etin chemotactic activities on neutrophils are regulated by PI-3K activationrdquo Journal of Leukocyte Biology vol 81 no 4 pp1093ndash1101 2007

[19] O Stoeltzing S A Ahmad W Liu et al ldquoAngiopoietin-1inhibits vascular permeability angiogenesis and growth of


hepatic colon cancer tumorsrdquo Cancer Research vol 63 no 12pp 3370ndash3377 2003

[20] I Cascone E Audero E Giraudo et al ldquoTie-2-dependentactivation of RhoA and Rac1 participates in endothelial cellmotility triggered by angiopoietin-1rdquo Blood vol 102 no 7 pp2482ndash2490 2003

[21] R Maliba S Lapointe P-E Neagoe A Brkovic and M GSirois ldquoAngiopoietins-1 and -2 are both capable of mediatingendothelial PAF synthesis intracellular signalling pathwaysrdquoCellular Signalling vol 18 no 11 pp 1947ndash1957 2006

[22] C Lemieux R Maliba J Favier J-F Theoret Y Merhi and MG Sirois ldquoAngiopoietins can directly activate endothelial cellsand neutrophils to promote proinflammatory responsesrdquoBloodvol 105 no 4 pp 1523ndash1530 2005

[23] S D McCarter P F H Lai R S Suen and D J StewartldquoRegulation of endothelin-1 by angiopoietin-1 implications forinflammationrdquo Experimental Biology andMedicine vol 231 no6 pp 985ndash991 2006

[24] S Dimmeler and A M Zeiher ldquoAkt takes center stage inangiogenesis signalingrdquo Circulation Research vol 86 no 1 pp4ndash5 2000

[25] R Harfouche H M Hassessian Y Guo et al ldquoMechanismswhich mediate the antiapoptotic effects of angiopoietin-1 onendothelial cellsrdquoMicrovascular Research vol 64 no 1 pp 135ndash147 2002

[26] C Daly V Wong E Burova et al ldquoAngiopoietin-1 modulatesendothelial cell function and gene expression via the transcrip-tion factor FKHR (FOXO1)rdquo Genes and Development vol 18no 9 pp 1060ndash1071 2004

[27] S Babaei K Teichert-Kuliszewska Q Zhang N Jones D JDumont andD J Stewart ldquoAngiogenic actions of angiopoietin-1 require endothelium-derived nitric oxiderdquo The AmericanJournal of Pathology vol 162 no 6 pp 1927ndash1936 2003

[28] J-X Chen M L Lawrence G Cunningham B W Christmanand B Meyrick ldquoHSP90 and Akt modulate Ang-1-inducedangiogenesis via NO in coronary artery endotheliumrdquo Journalof Applied Physiology vol 96 no 2 pp 612ndash620 2004

[29] R Maliba A Brkovic P-E Neagoe L R Villeneuve andM G Sirois ldquoAngiopoietin-mediated endothelial P-selectintranslocation cell signaling mechanismsrdquo Journal of LeukocyteBiology vol 83 no 2 pp 352ndash360 2008

[30] N Jones Z Master J Jones et al ldquoIdentification of TekTie2binding partners Binding to a multifunctional docking sitemediates cell survival and migrationrdquo Journal of BiologicalChemistry vol 274 no 43 pp 30896ndash30905 1999

[31] U Fiedler T Krissl S Koidl et al ldquoAngiopoietin-1 andangiopoietin-2 share the same binding domains in the Tie-2 receptor involving the first Ig-like loop and the epidermalgrowth factor-like repeatsrdquo Journal of Biological Chemistry vol278 no 3 pp 1721ndash1727 2003

[32] K G Peters C D Kontos P C Lin et al ldquoFunctionalsignificance of Tie2 signaling in the adult vasculaturerdquo RecentProgress in Hormone Research vol 59 pp 51ndash71 2004

[33] A Hashiramoto C Sakai K Yoshida et al ldquoAngiopoietin1 directly induces destruction of the rheumatoid joint bycooperative but independent signaling via ERKMAPK andphosphatidylinositol 3-kinaseAktrdquo Arthritis and Rheumatismvol 56 no 7 pp 2170ndash2179 2007

[34] J Gavard V Patel and J S Gutkind ldquoAngiopoietin-1 preventsVEGF-induced endothelial permeability by sequestering Src

through mDiardquo Developmental Cell vol 14 no 1 pp 25ndash362008

[35] Z Master N Jones J Tran J Jones R S Kerbel and DJ Dumont ldquoDok-R plays a pivotal role in angiopoietin-1-dependent cell migration through recruitment and activationof PakrdquoThe EMBO Journal vol 20 no 21 pp 5919ndash5928 2001

[36] D P Hughes M B Marron and N P J Brindle ldquoTheantiinflammatory endothelial tyrosine kinase Tie2 interactswith a novel nuclear factor-120581B inhibitor ABIN-2rdquo CirculationResearch vol 92 no 6 pp 630ndash636 2003

[37] A Tadros D P Hughes B J Dunmore and N P J BrindleldquoABIN-2 protects endothelial cells from death and has a role inthe antiapoptotic effect of angiopoietin-1rdquoBlood vol 102 no 13pp 4407ndash4409 2003

[38] E I Korpelainen M Karkkainen Y Gunji M Vikkula andK Alitalo ldquoEndothelial receptor tyrosine kinases activate theSTAT signaling pathway mutant Tie-2 causing venous malfor-mations signals a distinct STAT activation responserdquoOncogenevol 18 no 1 pp 1ndash8 1999

[39] C C Weber H Cai M Ehrbar et al ldquoEffects of protein andgene transfer of the angiopoietin-1 fibrinogen-like receptor-binding domain on endothelial and vessel organizationrdquo Journalof Biological Chemistry vol 280 no 23 pp 22445ndash22453 2005

[40] MThomas M Felcht K Kruse et al ldquoAngiopoietin-2 stimula-tion of endothelial cells induces alphavbeta3 integrin internal-ization and degradationrdquo The Journal of Biological Chemistryvol 285 no 31 pp 23842ndash23849 2010

[41] T Li Z Liu K Jiang and Q Ruan ldquoAngiopoietin2 enhancesdoxorubin resistance in HepG2 cells by upregulating survivinand Ref-1 via MSK1 activationrdquo Cancer Letters vol 337 no 2pp 276ndash284 2013

[42] T Etoh H Inoue S Tanaka G F Barnard S Kitano andM Mori ldquoAngiopoietin-2 is related to tumor angiogenesis ingastric carcinoma possible in vivo regulation via induction ofproteasesrdquo Cancer Research vol 61 no 5 pp 2145ndash2153 2001

[43] J Kosacka M Figiel J Engele H Hilbig M Majewski and KSpanel-Borowski ldquoAngiopoietin-1 promotes neurite outgrowthfrom dorsal root ganglion cells positive for Tie-2 receptorrdquo Celland Tissue Research vol 320 no 1 pp 11ndash19 2005

[44] O-H Lee J Xu J Fueyo et al ldquoExpression of the receptortyrosine kinase Tie2 in neoplastic glial cells is associated withintegrin 1205731-dependent adhesion to the extracellular matrixrdquoMolecular Cancer Research vol 4 no 12 pp 915ndash926 2006

[45] L M DeBusk Y Chen T Nishishita J Chen J W Thomasand P C Lin ldquoTie2 receptor tyrosine kinase a major mediatorof tumor necrosis factor 120572-induced angiogenesis in rheumatoidarthritisrdquo Arthritis and Rheumatism vol 48 no 9 pp 2461ndash2471 2003

[46] H Kanazawa S Nomura and K Asai ldquoRoles of angiopoietin-1 and angiopoietin-2 on airway microvascular permeability inasthmatic patientsrdquo Chest vol 131 no 4 pp 1035ndash1041 2007

[47] V Nanjappa R Raju B Muthusamy et al ldquoA comprehensivecurated reaction map of leptin signaling pathwayrdquo Journal ofProteomics and Bioinformatics vol 4 no 9 pp 184ndash189 2011

[48] M Bhattacharjee R Raju A Radhakrishnan et al ldquoA bioinfor-matics resource for TWEAK-Fn14 signaling pathwayrdquo Journalof Signal Transduction vol 2012 Article ID 376470 10 pages2012

[49] K Kandasamy S Keerthikumar R Raju et al ldquoPathBuildermdashopen source software for annotating and developing pathwayresourcesrdquo Bioinformatics vol 25 no 21 pp 2860ndash2862 2009


[50] M P van Iersel T Kelder A R Pico et al ldquoPresenting andexploring biological pathways with PathVisiordquo BMC Bioinfor-matics vol 9 article 399 2008

[51] R Raju V Nanjappa L Balakrishnan et al ldquoNetSlim high-confidence curated signaling mapsrdquoDatabase vol 2011 ArticleID bar032 2011

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Microbiology


Angiopoietin-4 (ANGPT4) is also known to be an agonist ofTIE2while angiopoietin-3 (ANGPT3) themouse ortholog ofangiopoietin-4 is reported to be antagonistic toTIE2 [11]Theother member of the TIE family is the orphan receptor TIE1It heterodimerizes with TIE2 and modulates TIE2 signalinginduced by ANGPT1 and ANGPT2 [12] ANGPT1 binding toTIE2 induces dissociation of the TIE1-TIE2 complex [12]Thissuggests that TIE2 signaling is regulated by the molecularbalance between ANGPT1 and ANGPT2 [6 13] and TIE1and TIE2 with another one being the ectodomain cleavageof TIE receptors [14] The activation of TIE2 is achievedby the assembly with tetrameric or higher order multimericangiopoietins clearly differentiating TIE2 from other tyro-sine kinase receptors [15] ANGPT1 induces the translocationof TIE2 to cell-cell junctions and transassociation in theform of homomeric complexes to activate the downstreamsignaling of TIE2 [16]

Binding of ANGPT1 to TIE2 leads to receptor dimer-ization and subsequent activation followed by autophospho-rylation at specific tyrosine residues [15 17] These phos-phorylated sites provide binding platform to a number ofeffector molecules to initiate downstream signaling cascadewhich ultimately controls various cellular responses includ-ing morphogenesis proliferation extracellular matrix inter-action permeability survival and differentiation [18ndash23]TIE2 interacts with p85 subunit of phosphatidylinositol-3-kinase (PI3K) via Tyr-1101 and activates PI3K-AKT pathwaywhich inhibits Smac release frommitochondria and increasesthe expression of survivin leading to survival and chemo-taxis of endothelial cells [18 24 25] AKT activation alsoinhibits forkhead transcription factor FKHR (FOXO1) whichprotects endothelial cells from apoptosis [26] ANGPT1 alsoinduces the PI3KAKT mediated activation of eNOS andNO release in endothelial cells [27 28] In endothelial cellsboth ANGPT1 and ANGPT2 also induce TIE2-dependenttranslocation of P-selectin through a PLCG1Ca2+ signalingpathway [29]

SH2 domain containing proteins such as growth factorreceptor-bound protein 2 (GRB2) growth factor receptor-bound protein 7 (GRB7) growth factor receptor-bound pro-tein 14 (GRB14) protein tyrosine phosphatase nonreceptortype 11 (SHP-2) and phosphoinositide-3-kinase (PI3K) isrecruited and transphosphorylated by TIE2 [30] GRB2 andSHC1 recruit SOS1 and lead to the activation of Ras-Raf-mitogen activated protein kinase (MAPK) pathway that reg-ulates platelet activating factor synthesis anti-inflammatoryresponses and endothelial cell migration proliferation per-meability and morphogenesis [5 20ndash22 31 32] ThroughSOS1 or PI3Ks angiopoietinTIE2 system also regulates theactivation of RAC1 RHOA CDC42 and focal adhesionkinase 1 to mediate cytoskeleton reorganization and migra-tion of endothelial and synovial cells [33] Angiopoietin-1 induced activation of RHOA results in sequestration ofSRC by DIAPH1 thereby preventing SRC association withVEGFR2 [34] Recruitment of dynamic complexes com-prising NCK adaptor protein 1 (NCK1) RAS p21 proteinactivator 1 (p120GAP) and P21 protein-activated kinase 1(PAK1) to TIE2 by the DOKs especially DOK2 has beenattributed to increased cell motility [35] TIE2 also interacts

with the inhibitor of nuclear factor kappa B (NF-kB) activityTNFAIP3 interacting protein 2 (ABIN-2) that inhibits NF-kB transcriptional activity and mediates anti-inflammatoryand antiapoptotic action [36 37] TIE2 activation inducesthe phosphorylation of STAT1 STAT3 and STAT5A5B andtheir subsequent translocation into nucleus to induce theexpression of the cell cycle inhibitor cyclin-dependent kinaseinhibitor 1A (p21) [38] ANGPT2 also interacts with integrinslike integrin 120572V1205735 120572V1205733 and 12057251205731 in endothelial cells withless affinity than TIE2 and can induce TIE2-independentsignaling [39] TIE2 also forms a complex with 12057251205733 andFAKANGPT2 induces phosphorylation of FAK at Serine91012057251205733 internalization and dissociation of p130CAS and talinfrom 12057251205733 [40] Recently ANGPT2 has also been shown toinduce the activation of ERKMSK1CREB pathway to impartcell survival and resistance to doxorubicin in HepG2 cells[41]

Besides the defects in vascular system and angiogene-sis [42ndash44] TIE2 signaling has also been associated withrheumatoid arthritis [45] and asthma [46] Considering theimportance of TIE2 signaling here we provide a manuallycurated enhanced network map of angiopoietin(s)-inducedTIE2-mediated signaling events as a reference platform forfurther biomedical investigations

2 Methods

We screened published research articles related to TIE2signaling NetPath criteria described earlier [47 48] werefollowed for the annotation of protein-protein interac-tions (PPIs) enzyme-substrate relationships and posttrans-lational modifications (PTMs) (catalytic events) Activa-tioninhibition status of proteins alterations in protein local-ization and also genes regulated at mRNA level by TIE2signaling were also documented PathBuilder an in-housepathway annotation tool was used for the curation of thesereactions [49] Each curated reaction was internally reviewedby trained biocurators followed by an external review by aPathway Authority an expert in the field (FMG coauthor ofthis paper)

3 Results and Discussion

Our analysis resulted in the cataloging of 140 uniquemolecules that are reported in TIE2 signaling Thesemolecules were part of 43 PPIs and 102 catalytic events23 activationinhibition events and 11 protein transloca-tion events We have also documented 124 and 65 genesthat were reported to be upregulated and downregulatedrespectively by TIE2 signaling in response to angiopoi-etin(s) in human cells The curated data for TIE2 sig-naling pathway is freely available to the scientific com-munity for visualization and download in different com-munity standard data exchange formats through NetPath[httpwwwnetpathorg] a resource of signaling path-ways These formats include Proteomics Standards Initiativefor Molecular Interaction (PSI-MI version 21) Biological


PAK1S204T423

STAT5BY699

STAT5AY694

AKT2S474

STAT5BY699

MAPK9T183 Y185

STAT5AY694

STAT3Y705

TEK

PIK3CA

BCAR1 P

PIK3R2P

GRB14 P

TEKY1108TEK Y1102 Y992Y1113

ANGPT1

ANGPT1

ANGPT1ANGPT1

CRK

RASA1

DOK4

Fibronectin

NCK1P

DOK2Y351


PIK3R1 P

GRB2

GRB7P

PTPN11P

ITGA5

PTK2Y861 S910

Y397

ITGB1

MAPK1Y187T185

MAPK3Y204T202MMP2

ILK


cell migration

RAC1GTP

NOS3S1179

TNIP2

PTPRB

ANGPT2

TEKP

CDKN1A


Oligomeric

MTORS2481

HIF1A

CXCL12



CBL

PLD2PLD1

RASGTP

RAF1P

MAP2K1S218

MAP2K2S222

NFKBIAP

Dissociation

RELA

RELA

CY

NU

Degradation

PDPK1P

PIP2

PIP3

ROCK1

CDC42GTP

RHOAGTP

DIAPH1



FOXO1S256T24S319

GJA5


endothelial cells

FOXO1S319T24

S256

CY

NU

SELP

PLCG1

SELP

PKC

Ca2+


NR4A1


PXN P

PLG

FESP

FYN P


RPS6KB1T389S411

EGR1

SOS1

ARHGAP5 P

MAPK14Y182T180

RPS6KA1S380

ELK1S383

MAP3K3S526

MAPK7 P



synthesis

PLA2G5

Phospholipids

Lyso-PAF

Dissociation

DLL4

CTNNB1

MAPK8Y185 T183

BCL2


FOXO3S253 S318S321

BIRC5



MAP2K4S80

STAT3 Y705

STAT1 Y701

SPHK1P


permeability


HSP90AA1

ITGB3ITGAV

Ub


and degradation

Dissociation


SHC1P

SRC

TIE1 P

BMXP


human MECs

ROS

NOX

NO


endothelial cells

TEK



MEF2A

MEF2C

MEF2D

EGR1



formation


Dissociation

TLN1

VEGFA

KLF2

CY

NU

EGR1

CY

NU

STAT1Y701

IL8

JUNY182T180


MAPK14T180Y182

ANGPT4

TEK TEK

ANGPT2 ANGPT4


ITGAVITGB5ITGAV


Preventsbinding of

SRC to VEGFR2

Dissociation

PLG

TIAM1

PRKCZ

CTNNB1ZAP70P

EGFRP

ITGB2 P

GNA11 P

PGK1 P

PDIA3 P

ALDOA P

GTPBP3 P

HAGHP

MASP2 P

ADSS P

TXNRD3P

TXNRD1P CTPS1 P

GYS1 P

LIMCH1P

ProteinReceptor

Protein

Protein

mRNA

Small molecule

Ligand

Induced activation

Enzyme complex

Inhibition

Autocatalysis

Transport




Translocation

Ubiquitination

PTM state


Dephosphorylation

Phosphorylation


CYPM

NU

Molecule

ANGPT2Inhibition of


ANGPT2expression

IKK






4 Conclusions




Acknowledgments


References































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


PAK1S204T423

STAT5BY699

STAT5AY694

AKT2S474

STAT5BY699

MAPK9T183 Y185

STAT5AY694

STAT3Y705

TEK

PIK3CA

BCAR1 P

PIK3R2P

GRB14 P

TEKY1108TEK Y1102 Y992Y1113

ANGPT1

ANGPT1

ANGPT1ANGPT1

CRK

RASA1

DOK4

Fibronectin

NCK1P

DOK2Y351


PIK3R1 P

GRB2

GRB7P

PTPN11P

ITGA5

PTK2Y861 S910

Y397

ITGB1

MAPK1Y187T185

MAPK3Y204T202MMP2

ILK


cell migration

RAC1GTP

NOS3S1179

TNIP2

PTPRB

ANGPT2

TEKP

CDKN1A


Oligomeric

MTORS2481

HIF1A

CXCL12



CBL

PLD2PLD1

RASGTP

RAF1P

MAP2K1S218

MAP2K2S222

NFKBIAP

Dissociation

RELA

RELA

CY

NU

Degradation

PDPK1P

PIP2

PIP3

ROCK1

CDC42GTP

RHOAGTP

DIAPH1



FOXO1S256T24S319

GJA5


endothelial cells

FOXO1S319T24

S256

CY

NU

SELP

PLCG1

SELP

PKC

Ca2+


NR4A1


PXN P

PLG

FESP

FYN P


RPS6KB1T389S411

EGR1

SOS1

ARHGAP5 P

MAPK14Y182T180

RPS6KA1S380

ELK1S383

MAP3K3S526

MAPK7 P



synthesis

PLA2G5

Phospholipids

Lyso-PAF

Dissociation

DLL4

CTNNB1

MAPK8Y185 T183

BCL2


FOXO3S253 S318S321

BIRC5



MAP2K4S80

STAT3 Y705

STAT1 Y701

SPHK1P


permeability


HSP90AA1

ITGB3ITGAV

Ub


and degradation

Dissociation


SHC1P

SRC

TIE1 P

BMXP


human MECs

ROS

NOX

NO


endothelial cells

TEK



MEF2A

MEF2C

MEF2D

EGR1



formation


Dissociation

TLN1

VEGFA

KLF2

CY

NU

EGR1

CY

NU

STAT1Y701

IL8

JUNY182T180


MAPK14T180Y182

ANGPT4

TEK TEK

ANGPT2 ANGPT4


ITGAVITGB5ITGAV


Preventsbinding of

SRC to VEGFR2

Dissociation

PLG

TIAM1

PRKCZ

CTNNB1ZAP70P

EGFRP

ITGB2 P

GNA11 P

PGK1 P

PDIA3 P

ALDOA P

GTPBP3 P

HAGHP

MASP2 P

ADSS P

TXNRD3P

TXNRD1P CTPS1 P

GYS1 P

LIMCH1P

ProteinReceptor

Protein

Protein

mRNA

Small molecule

Ligand

Induced activation

Enzyme complex

Inhibition

Autocatalysis

Transport




Translocation

Ubiquitination

PTM state


Dephosphorylation

Phosphorylation


CYPM

NU

Molecule

ANGPT2Inhibition of


ANGPT2expression

IKK






4 Conclusions




Acknowledgments


References































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




4 Conclusions




Acknowledgments


References































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology












































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

research article signaling network map of endothelial tek … · 2019. 7. 31. · tie interacts...

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