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/ NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions 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. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions in vitro In vitro phosphotransferase phosphotransferase assay phosphotransferase weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions 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. pull down pull down anti tag western anti tag western blot anti tag western Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis Additional details on source of TGFBR1 supplied by the author. hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis Nucleoside diphosphate kinase activity assayed by the formation of GTP from GDP hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions human-hep_3b Homo sapiens hepatocellular carcinoma (HCC) cells (p53-null). hep_3b human hepatocellular carcinoma (HCC) cells (p53-null). anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis Additional details on source of PDPK1 and SGK1 supplied by author. hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions 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. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions 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. anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions human-hep_3b Homo sapiens hepatocellular carcinoma (HCC) cells (p53-null). hep_3b human hepatocellular carcinoma (HCC) cells (p53-null). confocal microscopy confocal microscopy immunostaining immunostaining Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. NM23-H1 tumor suppressor physically interacts with serine-threonine kinase receptor-associated protein, a transforming growth factor-beta (TGF-beta) receptor-interacting protein, and negatively regulates TGF-beta signaling. J. Biol. Chem. (0021-9258) 2007 imex curation Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. hyunha@chungbuk.ac.kr 09-April-2009: Contacted by JYOTI. Human HeLa cDNA davet Only protein-protein interactions human-sk_n_be(2) Human caucasian neuroblastoma cell line sk_n_be(2) Human neuroblastoma cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Seong HA., Jung H., Ha H. Apoptosis - Interactions involving proteins with a function related to apoptosis hyunha@chungbuk.ac.kr Accepted 2008-Not a month!-29 by DAVET. 09-April-2009: Contacted by JYOTI. Human HeLa cDNA J. Biol. Chem. (0021-9258) 2007 davet Only protein-protein interactions imex curation ndkb_human Nucleoside diphosphate kinase B nm23-H2 C-myc purine-binding transcription factor PUF NME2 NM23B Histidine protein kinase NDKB protein protein human Homo sapiens Human MANLERTFIAIKPDGVQRGLVGEIIKRFEQKGFRLVAMKFLRASEEHLKQHYIDLKDRPFFPGLVKYMNSGPVVAMVWEGLNVVKTGRVMLGETNPADSKPGTIRGDFCIQVGRNIIHGSDSVKSAEKEISLWFKPEELVDYKSCAHDWVYE DIP protein P22392 original sequence version: 182 1A5C3F84D7AD272C ndka_human Nucleoside diphosphate kinase A Tumor metastatic process-associated protein Metastasis inhibition factor nm23 Granzyme A-activated DNase NME1 NDPKA NM23 NM23-H1 protein protein human Homo sapiens Human MANCERTFIAIKPDGVQRGLVGEIIKRFEQKGFRLVGLKFMQASEDLLKEHYVDLKDRPFFAGLVKYMHSGPVVAMVWEGLNVVKTGRVMLGETNPADSKPGTIRGDFCIQVGRNIIHGSDSVESAEKEIGLWFHPEELVDYTSCAQNWIYE AAE9C0DF63CB70A1 sgk1_human Serine/threonine-protein kinase Sgk1 SGK1 SGK Serum/glucocorticoid-regulated kinase 1 protein protein human Homo sapiens Human MTVKTEAAKGTLTYSRMRGMVAILIAFMKQRRMGLNDFIQKIANNSYACKHPEVQSILKISQPQEPELMNANPSPPPSPSQQINLGPSSNPHAKPSDFHFLKVIGKGSFGKVLLARHKAEEVFYAVKVLQKKAILKKKEEKHIMSERNVLLKNVKHPFLVGLHFSFQTADKLYFVLDYINGGELFYHLQRERCFLEPRARFYAAEIASALGYLHSLNIVYRDLKPENILLDSQGHIVLTDFGLCKENIEHNSTTSTFCGTPEYLAPEVLHKQPYDRTVDWWCLGAVLYEMLYGLPPFYSRNTAEMYDNILNKPLQLKPNITNSARHLLEGLLQKDRTKRLGAKDDFMEIKSHVFFSLINWDDLINKKITPPFNPNVSGPNDLRHFDPEFTEEPVPNSIGKSPDSVLVTASVKEAAEAFLGFSYAPPTDSFL DIP protein O00141 original sequence version: 145 F3697C63AB1F499D gdp guanosine 5-diphosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) tgfr1_human TGF-beta receptor type-1 TGFBR1 Transforming growth factor-beta receptor type I TGF-beta type I receptor Serine/threonine-protein kinase receptor R4 Activin receptor-like kinase 5 ALK5 SKR4 Activin A receptor type II-like protein kinase of 53kD protein protein human Homo sapiens Human MEAAVAAPRPRLLLLVLAAAAAAAAALLPGATALQCFCHLCTKDNFTCVTDGLCFVSVTETTDKVIHNSMCIAEIDLIPRDRPFVCAPSSKTGSVTTTYCCNQDHCNKIELPTTVKSSPGLGPVELAAVIAGPVCFVCISLMLMVYICHNRTVIHHRVPNEEDPSLDRPFISEGTTLKDLIYDMTTSGSGSGLPLLVQRTIARTIVLQESIGKGRFGEVWRGKWRGEEVAVKIFSSREERSWFREAEIYQTVMLRHENILGFIAADNKDNGTWTQLWLVSDYHEHGSLFDYLNRYTVTVEGMIKLALSTASGLAHLHMEIVGTQGKPAIAHRDLKSKNILVKKNGTCCIADLGLAVRHDSATDTIDIAPNHRVGTKRYMAPEVLDDSINMKHFESFKRADIYAMGLVFWEIARRCSIGGIHEDYQLPYYDLVPSDPSVEEMRKVVCEQKLRPNIPNRWQSCEALRVMAKIMRECWYANGAARLTALRIKKTLSQLSQQEGIKM DIP protein P36897 original sequence version: 161 179F11404725DDCB pdpk1_human 3-phosphoinositide-dependent protein kinase 1 PDPK1 PDK1 protein protein human Homo sapiens Human MARTTSQLYDAVPIQSSVVLCSCPSPSMVRTQTESSTPPGIPGGSRQGPAMDGTAAEPRPGAGSLQHAQPPPQPRKKRPEDFKFGKILGEGSFSTVVLARELATSREYAIKILEKRHIIKENKVPYVTRERDVMSRLDHPFFVKLYFTFQDDEKLYFGLSYAKNGELLKYIRKIGSFDETCTRFYTAEIVSALEYLHGKGIIHRDLKPENILLNEDMHIQITDFGTAKVLSPESKQARANSFVGTAQYVSPELLTEKSACKSSDLWALGCIIYQLVAGLPPFRAGNEYLIFQKIIKLEYDFPEKFFPKARDLVEKLLVLDATKRLGCEEMEGYGPLKAHPFFESVTWENLHQQTPPKLTAYLPAMSEDDEDCYGNYDNLLSQFGCMQVSSSSSSHSLSASDTGLPQRSGSNIEQYIHDLDSNSFELDLQFSEDEKRLLLEKQAGGNPWHQFVENNLILKMGPVDKRKGLFARRRQLLLTEGPHLYYVDPVNKVLKGEIPWSQELRPEAKNFKTFFVHTPNRTYYLMDPSGNAHKWCRKIQEVWRQRYQSHPDAAVQ DIP protein O15530 original sequence version: 169 DIP protein O15530-1 original sequence version: 178 ED8C0306DC4D0653 strap_human Serine-threonine kinase receptor-associated protein UNR-interacting protein WD-40 repeat protein PT-WD MAP activator with WD repeats STRAP MAWD UNRIP protein protein human Homo sapiens Human MAMRQTPLTCSGHTRPVVDLAFSGITPYGYFLISACKDGKPMLRQGDTGDWIGTFLGHKGAVWGATLNKDATKAATAAADFTAKVWDAVSGDELMTLAHKHIVKTVDFTQDSNYLLTGGQDKLLRIYDLNKPEAEPKEISGHTSGIKKALWCSEDKQILSADDKTVRLWDHATMTEVKSLNFNMSVSSMEYIPEGEILVITYGRSIAFHSAVSLDPIKSFEAPATINSASLHPEKEFLVAGGEDFKLYKYDYNSGEELESYKGHFGPIHCVRFSPDGELYASGSEDGTLRLWQTVVGKTYGLWKCVLPEEDSGELAKPKIGFPETTEEELEEIASENSDCIFPSAPDVKA CFCB34D3946290E2 strap-nme1 1 14 unspecified role unspecified role bait bait 19 unspecified role unspecified role prey prey physical association physical association Previously, a screen for NM23-H1-interacting proteins that could potentially modulate its activity identified serine-threonine kinase receptor-associated protein (STRAP), a transforming growth factor (TGF)-beta receptor-interacting protein. strap-nme1-1 4 19 unspecified role unspecified role prey prey 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 Q9Y3F4:p.Cys152Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 17:00:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys152ser' Tue Jun 30 18:24:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys152Ser' Q9Y3F4:p.Cys270Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 19:57:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys270ser' Tue Jun 30 15:39:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys270Ser' Q9Y3F4:p.[Cys152Ser;Cys270Ser] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Fri Jul 22 10:53:35 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'cys270ser,cys152ser' Wed May 02 16:35:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys152ser,cys270ser' Tue Jun 30 19:18:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys152Ser;Cys270Ser]' 14 unspecified role unspecified role bait bait 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 P15531:p.Cys145Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 16:24:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys145ser' Tue Jun 30 18:01:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys145Ser' physical association physical association FLAG-STRAP was detected in association with NM23-H1 only when coexpressed with GST-NM23-H1 (Fig. 1A, top). P15531; Q9UQC8 transfection Figs. 1A, 2A, 2B strap-nme1-2 2 19 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey physical association physical association NM23-H1 was present in anti-STRAP immune complexes from all cell lines examined, including 293T, Hep3B, and SK-N-BE(2)C (18) cells (Fig. 1B), demonstrating that NM23-H1 physically interacts with STRAP in vivo. Fig. 1B strap-nme1-3 7 14 unspecified role unspecified role prey prey 19 unspecified role unspecified role bait bait physical association physical association NM23-H1 was present in anti-STRAP immune complexes from all cell lines examined, including 293T, Hep3B, and SK-N-BE(2)C (18) cells (Fig. 1B), demonstrating that NM23-H1 physically interacts with STRAP in vivo. Fig. 1B strap-nme1-4 12 19 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey physical association physical association NM23-H1 was present in anti-STRAP immune complexes from all cell lines examined, including 293T, Hep3B, and SK-N-BE(2)C (18) cells (Fig. 1B), demonstrating that NM23-H1 physically interacts with STRAP in vivo. Fig. 1B strap-nme1-5 11 14 unspecified role unspecified role 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 19 unspecified role unspecified role 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 colocalization colocalization Both STRAP and NM23-H1 were distributed mainly in the cytoplasm and colocalized with each other, as seen in the merged image in Fig. 1C. Fig. 1C strap-nme1-6 10 19 unspecified role unspecified role prey prey 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 14 unspecified role unspecified role bait bait physical association physical association To determine whether the interaction of NM23-H1 and STRAP was redox-dependent, we examined complex formation in 293T cells transiently expressing FLAG-STRAP using in vivo binding assays. Fig. 2C Reducing conditions - dithiothreitol Reducing conditions - beta-mercaptoethanol strap-nme1-7 6 14 unspecified role unspecified role neutral component neutral component region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 19 unspecified role unspecified role neutral component neutral component Q9Y3F4:p.[Cys152Ser;Cys270Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:44:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys152ser,cys270ser' Tue Jun 30 17:00:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys152Ser;Cys270Ser]' ecoli Escherichia coli (strain K12) direct interaction direct interaction As shown in Fig. 2D, a shift in the mobility of autophosphorylated NM23-H1 was clearly evident upon incubation in the presence of wild-type STRAP Fig. 2D strap-nme1-8 9 19 unspecified role unspecified role prey prey 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 Q9Y3F4:p.Cys152Ser mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 18:16:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys152ser' Tue Jun 30 18:08:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys152Ser' Q9Y3F4:p.Cys270Ser mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 20:22:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys270ser' Tue Jun 30 15:54:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys270Ser' wd4 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 14 unspecified role unspecified role bait bait 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 physical association physical association To map the domain(s) within STRAP required for its association with NM23-H1, we generated a set of eight STRAP deletion mutants (Fig. 3A) and examined their ability to interact with NM23-H1 in an in vivo binding assay. P15531; Q9UQC8 transfection Figs. 3 nme1-1 4 14 unspecified role unspecified role prey prey 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 14 unspecified role unspecified role bait bait 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 physical association physical association The expression of the NM23-H1 mutants NM23-H1(C4S), NM23-H1(C109S), and NM23-H1(C145S) had no effect on complex formation between NM23-H1 and NM23-H1 (Fig. 4A) Fig. 4A nme1-nme2-1 4 14 unspecified role unspecified role prey prey 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 13 unspecified role unspecified role bait bait 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 physical association physical association The expression of the NM23-H1 mutants NM23-H1(C4S), NM23-H1(C109S), and NM23-H1(C145S) had no effect on complex formation between NM23-H1 and NM23-H1 (Fig. 4A) or NM23-H1 and NM23-H2 (Fig. 4B), Fig. 4B strap-9 4 19 unspecified role unspecified role prey prey 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 19 unspecified role unspecified role bait bait 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 physical association physical association As shown in Fig. 4F, no changes in the self-association of STRAP were detected in the presence of the STRAP mutants. Fig. 4F nme1-2 8 14 putative self putative self putative self putative self 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 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. phosphorylation phosphorylation reaction phosphorylation true To determine whether the Cys -> Ser mutations of NM23-H1 affected its biochemical properties, we examined the autophosphorylation (Fig. 5B) Fig. 5B Autophosphorylation of NME1 nme1-3 3 14 phosphate donor phosphate donor neutral component neutral component 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 region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 14 phosphate acceptor phosphate acceptor neutral component neutral component 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 ecoli Escherichia coli (strain K12) phosphotransfer phosphotransfer reaction phosphotransfer Phosphotransferase assays were performed with the indicated wild type (WT) and NM23-H1 mutants Fig. 5C nme1-gdp 5 14 enzyme enzyme neutral component neutral component 16 enzyme target enzyme target substrate substrate neutral component neutral component phosphotransfer phosphotransfer reaction phosphotransfer NDP kinase activity was assayed in the presence of purified wild-type and mutant NM23-H1 proteins Fig. 5D strap-tgfbr1-1 4 19 unspecified role unspecified role prey prey 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 17 unspecified role unspecified role bait bait 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 physical association physical association 293T cells were transfected with expression vectors for wild-type and mutant forms of STRAP, together with a GST fusion protein of a constitutively active form of the TGF-beta type 1 receptor, GST-TbetaR1(TD). Cell lysates were incubated with glutathione-Sepharose beads, and protein complexes were analyzed by immunoblot using an anti-FLAG antibody. P36897; Q9UQC8 transfection Figs. 4G pdpk1-sgk-1 8 15 enzyme target enzyme target substrate substrate neutral component neutral component 18 enzyme enzyme neutral component neutral component 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. phosphorylation phosphorylation reaction phosphorylation PDK1 was immunoprecipitated from the transfected cells, and its activity was measured in an in vitro kinase assay using serum- and glucocorticoid-inducible kinase as a substrate (31). Fig. 5A Coexpression of STRAP mutants with PDK1 prior to purification resulted in a significant increase in PDK1 activity.