IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Cancer Cell (1535-6108) 2009 imex curation Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 11 human-mcf10a Human non-transformed breast epithelial cell line mcf10a non-transformed human breast epithelial cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org Accepted 2010-MAR-12 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 11 imex curation Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Cancer Cell (1535-6108) 2009 imex curation Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 11 in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org Accepted 2010-MAR-12 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 11 imex curation Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Cancer Cell (1535-6108) 2009 imex curation Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 11 human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference pull down pull down de novo protein sequence de novo protein sequencing by mass spectrometry de novo protein sequence Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org Accepted 2010-MAR-12 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 11 imex curation Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Cancer Cell (1535-6108) 2009 imex curation Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 11 human-mcf10a Human non-transformed breast epithelial cell line mcf10a non-transformed human breast epithelial cell line pull down pull down western blot western blot Immuno blot Immuno blot Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org Accepted 2010-MAR-12 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 11 imex curation Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Cancer Cell (1535-6108) 2009 imex curation Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 11 human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org Accepted 2010-MAR-12 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 11 imex curation Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Rak functions as a tumor suppressor by regulating PTEN protein stability and function. Cancer Cell (1535-6108) 2009 imex curation Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 11 human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yim EK., Peng G., Dai H., Hu R., Li K., Lu Y., Mills GB., Meric-Bernstam F., Hennessy BT., Craven RJ., Lin SY. Apoptosis - Interactions involving proteins with a function related to apoptosis U2OS cells were transiently transfected with empty vector or FLAG-PTEN plasmids. Whole cellular extracts were immunoprecipitated with anti-FLAG M2 affinity gel. Bead-bound immunocomplexes were eluted with 3 � FLAG peptide and subjected to SDS-PAGE. Silver staining was performed with a SilverSNAP mass spectrometry kit. Specific bands were excised and digested, and the peptides were analyzed by mass spectrometry. Source of flag-pten supplied by author. sylin@mdanderson.org Accepted 2010-MAR-12 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 11 imex curation pten_human Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN Phosphatase and tensin homolog Mutated in multiple advanced cancers 1 PTEN MMAC1 TEP1 protein protein human Homo sapiens Human MTAIIKEIVSRNKRRYQEDGFDLDLTYIYPNIIAMGFPAERLEGVYRNNIDDVVRFLDSKHKNHYKIYNLCAERHYDTAKFNCRVAQYPFEDHNPPQLELIKPFCEDLDQWLSEDDNHVAAIHCKAGKGRTGVMICAYLLHRGKFLKAQEALDFYGEVRTRDKKGVTIPSQRRYVYYYSYLLKNHLDYRPVALLFHKMMFETIPMFSGGTCNPQFVVCQLKVKIYSSNSGPTRREDKFMYFEFPQPLPVCGDIKVEFFHKQNKMLKKDKMFHFWVNTFFIPGPEETSEKVENGSLCDQEIDSICSIERADNDKEYLVLTLTKNDLDKANKDKANRYFSPNFKVKLYFTKTVEEPSNPEASSSTSVTPDVSDNEPDHYRYSDTTDSDPENEPFDEDQHTQITKV 75F97C3DD6802BA9 nedd4_human E3 ubiquitin-protein ligase NEDD4 Neural precursor cell expressed developmentally down-regulated protein 4 NEDD4 KIAA0093 NEDD4-1 Cell proliferation-inducing gene 53 protein PIG53 HECT-type E3 ubiquitin transferase NEDD4 protein protein human Homo sapiens Human MAQSLRLHFAARRSNTYPLSETSGDDLDSHVHMCFKRPTRISTSNVVQMKLTPRQTALAPLIKENVQSQERSSVPSSENVNKKSSCLQISLQPTRYSGYLQSSNVLADSDDASFTCILKDGIYSSAVVDNELNAVNDGHLVSSPAICSGSLSNFSTSDNGSYSSNGSDFGSCASITSGGSYTNSVISDSSSYTFPPSDDTFLGGNLPSDSTSNRSVPNRNTTPCEIFSRSTSTDPFVQDDLEHGLEIMKLPVSRNTKIPLKRYSSLVIFPRSPSTTRPTSPTSLCTLLSKGSYQTSHQFIISPSEIAHNEDGTSAKGFLSTAVNGLRLSKTICTPGEVRDIRPLHRKGSLQKKIVLSNNTPRQTVCEKSSEGYSCVSVHFTQRKAATLDCETTNGDCKPEMSEIKLNSDSEYIKLMHRTSACLPSSQNVDCQININGELERPHSQMNKNHGILRRSISLGGAYPNISCLSSLKHNCSKGGPSQLLIKFASGNEGKVDNLSRDSNRDCTNELSNSCKTRDDFLGQVDVPLYPLPTENPRLERPYTFKDFVLHPRSHKSRVKGYLRLKMTYLPKTSGSEDDNAEQAEELEPGWVVLDQPDAACHLQQQQEPSPLPPGWEERQDILGRTYYVNHESRRTQWKRPTPQDNLTDAENGNIQLQAQRAFTTRRQISEETESVDNRESSENWEIIREDEATMYSNQAFPSPPPSSNLDVPTHLAEELNARLTIFGNSAVSQPASSSNHSSRRGSLQAYTFEEQPTLPVLLPTSSGLPPGWEEKQDERGRSYYVDHNSRTTTWTKPTVQATVETSQLTSSQSSAGPQSQASTSDSGQQVTQPSEIEQGFLPKGWEVRHAPNGRPFFIDHNTKTTTWEDPRLKIPAHLRGKTSLDTSNDLGPLPPGWEERTHTDGRIFYINHNIKRTQWEDPRLENVAITGPAVPYSRDYKRKYEFFRRKLKKQNDIPNKFEMKLRRATVLEDSYRRIMGVKRADFLKARLWIEFDGEKGLDYGGVAREWFFLISKEMFNPYYGLFEYSATDNYTLQINPNSGLCNEDHLSYFKFIGRVAGMAVYHGKLLDGFFIRPFYKMMLHKPITLHDMESVDSEYYNSLRWILENDPTELDLRFIIDEELFGQTHQHELKNGGSEIVVTNKNKKEYIYLVIQWRFVNRIQKQMAAFKEGFFELIPQDLIKIFDENELELLMCGLGDVDVNDWREHTKYKNGYSANHQVIQWFWKAVLMMDSEKRIRLLQFVTGTSRVPMNGFAELYGSNGPQSFTVEQWGTPEKLPRAHTCFNRLDLPPYESFEELWDKLQMAIENTQGFDGVD DIP protein P46934 original sequence version: 76 D56EBBC50A34F13B frk_human Tyrosine-protein kinase FRK PTK5 RAK FYN-related kinase Nuclear tyrosine protein kinase RAK FRK Protein-tyrosine kinase 5 protein protein human Homo sapiens Human MSNICQRLWEYLEPYLPCLSTEADKSTVIENPGALCSPQSQRHGHYFVALFDYQARTAEDLSFRAGDKLQVLDTLHEGWWFARHLEKRRDGSSQQLQGYIPSNYVAEDRSLQAEPWFFGAIGRSDAEKQLLYSENKTGSFLIRESESQKGEFSLSVLDGAVVKHYRIKRLDEGGFFLTRRRIFSTLNEFVSHYTKTSDGLCVKLGKPCLKIQVPAPFDLSYKTVDQWEIDRNSIQLLKRLGSGQFGEVWEGLWNNTTPVAVKTLKPGSMDPNDFLREAQIMKNLRHPKLIQLYAVCTLEDPIYIITELMRHGSLQEYLQNDTGSKIHLTQQVDMAAQVASGMAYLESRNYIHRDLAARNVLVGEHNIYKVADFGLARVFKVDNEDIYESRHEIKLPVKWTAPEAIRSNKFSIKSDVWSFGILLYEIITYGKMPYSGMTGAQVIQMLAQNYRLPQPSNCPQQFYNIMLECWNAEPKERPTFETLRWKLEDYFETDSSYSDANNFIR 06EC050DDBCD930B pten-frk 3 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association We demonstrated that Rak tyrosine kinase represents one of the major PTEN-associated proteins. Fig1A. pten-frk-1 6 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association To validate the mass spectrometry result, we performed immunoprecipitation/western blot analysis and found that PTEN coprecipitated with Rak. Fig 1B. pten-frk-2 6 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association To validate the mass spectrometry result, we performed immunoprecipitation/western blot analysis and found that PTEN coprecipitated with Rak. Fig 1B. pten-frk-3 5 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait sh3 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association PTEN interacts with the Rak-SH3 domain. Fig 1C. The SH3 domain coodinates were inferred from the UniProt entry. pten-frk-4 5 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c2 domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain phosp_domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association The binding of Rak and PTEN is mediated through the interaction between the SH3 domain of Rak and the C2 domain (and perhaps the phosphatase domain as well) of PTEN. Fig 1D. pten-nedd 1 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Rak knockdown in MCF10A cells resulted in a substantial increase in the association between endogenous PTEN and NEDD4-1. Fig 2D. The interaction between NEDD4 and PTEN is markedly increased in the absence of Rak. pten-nedd-1 1 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association Rak knockdown in MCF10A cells resulted in a substantial increase in the association between endogenous PTEN and NEDD4-1. Fig 2D. The interaction between NEDD4 and PTEN is markedly increased in the absence of Rak. pten-frk-6 2 9 enzyme enzyme neutral component neutral component region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells 7 enzyme target enzyme target substrate substrate neutral component neutral component c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecobb Escherichia coli (strain B / BL21) phosphorylation phosphorylation reaction phosphorylation In an in vitro kinase assay Rak could induce PTEN phosphorylation. Fig 3A. pten-frk-7 2 7 enzyme target enzyme target substrate substrate neutral component neutral component region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P60484:p.Tyr336Phe Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 17:17:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr336phe' Tue Jun 30 18:40:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr336Phe' human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 9 enzyme enzyme neutral component neutral component P42685:p.Lys262Arg Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain K R Wed May 02 20:00:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys262arg' Tue Jun 30 15:49:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys262Arg' region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobd Escherichia coli (strain B / BL21-DE3) phosphorylation phosphorylation reaction phosphorylation The mutation of Tyr366 abolished the phosphorylation of PTEN by Rak. A Kinase dead mutant (K262R) was unable to phosphorylate PTEN. Fig 3C. pten-nedd4 5 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P60484:p.Tyr336Phe mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 20:10:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr336phe' Tue Jun 30 20:06:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr336Phe' physical association physical association Fig 3F. The E3 ligase NEDD4-1 has higher affinity to the Y336F mutant than wild type TPEN. Authors confirm that Fig 3F used U2OS cells. pten-ubi-2 4 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association PTEN polyubiquitination was significantly decreased in the absence of NEDD4-1. Fig 2E.