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/ c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. Nature communications 2017 imex curation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. liqinxi@xmu.edu.cn 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions human-hct116 Human colon carcinoma HCT116 cells hct116 Human colon carcinoma line HCT116 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. waiting to send out pending ability to add GO XRefs to Kinases assays liqinxi@xmu.edu.cn Accepted 2017-JUN-20 AT 15:42 BST AT 15:42 BST by MGT Nature communications 2017 imex curation 2G and 2I are not IPs - they are part of the evidence for the phosphorylated residue, which is why I added them since, apart from not blotting src they are identical to 2A/B etc. Delete if it really upsets you. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. Nature communications 2017 imex curation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. liqinxi@xmu.edu.cn 14-Jul-2017: Contacted by IntAct-Help. 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 Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. waiting to send out pending ability to add GO XRefs to Kinases assays liqinxi@xmu.edu.cn Accepted 2017-JUN-20 AT 15:42 BST AT 15:42 BST by MGT Nature communications 2017 imex curation 2G and 2I are not IPs - they are part of the evidence for the phosphorylated residue, which is why I added them since, apart from not blotting src they are identical to 2A/B etc. Delete if it really upsets you. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. Nature communications 2017 imex curation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. liqinxi@xmu.edu.cn 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. waiting to send out pending ability to add GO XRefs to Kinases assays liqinxi@xmu.edu.cn Accepted 2017-JUN-20 AT 15:42 BST AT 15:42 BST by MGT Nature communications 2017 imex curation 2G and 2I are not IPs - they are part of the evidence for the phosphorylated residue, which is why I added them since, apart from not blotting src they are identical to 2A/B etc. Delete if it really upsets you. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. Nature communications 2017 imex curation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. liqinxi@xmu.edu.cn 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay anti tag western anti tag western blot anti tag western Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. waiting to send out pending ability to add GO XRefs to Kinases assays liqinxi@xmu.edu.cn Accepted 2017-JUN-20 AT 15:42 BST AT 15:42 BST by MGT Nature communications 2017 imex curation 2G and 2I are not IPs - they are part of the evidence for the phosphorylated residue, which is why I added them since, apart from not blotting src they are identical to 2A/B etc. Delete if it really upsets you. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. Nature communications 2017 imex curation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. liqinxi@xmu.edu.cn 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down tag visualisation tag visualisation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. waiting to send out pending ability to add GO XRefs to Kinases assays liqinxi@xmu.edu.cn Accepted 2017-JUN-20 AT 15:42 BST AT 15:42 BST by MGT Nature communications 2017 imex curation 2G and 2I are not IPs - they are part of the evidence for the phosphorylated residue, which is why I added them since, apart from not blotting src they are identical to 2A/B etc. Delete if it really upsets you. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. Nature communications 2017 imex curation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. liqinxi@xmu.edu.cn 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. waiting to send out pending ability to add GO XRefs to Kinases assays crosslinker disuccinimidyl suberate (DSS) liqinxi@xmu.edu.cn Accepted 2017-JUN-20 AT 15:42 BST AT 15:42 BST by MGT Nature communications 2017 imex curation 2G and 2I are not IPs - they are part of the evidence for the phosphorylated residue, which is why I added them since, apart from not blotting src they are identical to 2A/B etc. Delete if it really upsets you. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. c-Src phosphorylation and activation of hexokinase promotes tumorigenesis and metastasis. Nature communications 2017 imex curation Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. liqinxi@xmu.edu.cn 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Zhang J., Wang S., Jiang B., Huang L., Ji Z., Li X., Zhou H., Han A., Chen A., Wu Y., Ma H., Zhao W., Zhao Q., Xie C., Sun X., Zhou Y., Huang H., Suleman M., Lin F., Zhou L., Tian F., Jin M., Cai Y., Zhang N., Li Q. waiting to send out pending ability to add GO XRefs to Kinases assays liqinxi@xmu.edu.cn Accepted 2017-JUN-20 AT 15:42 BST AT 15:42 BST by MGT Nature communications 2017 imex curation 2G and 2I are not IPs - they are part of the evidence for the phosphorylated residue, which is why I added them since, apart from not blotting src they are identical to 2A/B etc. Delete if it really upsets you. hxk2_human Hexokinase-2 Hexokinase type II Muscle form hexokinase HK2 Hexokinase-B protein protein human Homo sapiens Human MIASHLLAYFFTELNHDQVQKVDQYLYHMRLSDETLLEISKRFRKEMEKGLGATTHPTAAVKMLPTFVRSTPDGTEHGEFLALDLGGTNFRVLWVKVTDNGLQKVEMENQIYAIPEDIMRGSGTQLFDHIAECLANFMDKLQIKDKKLPLGFTFSFPCHQTKLDESFLVSWTKGFKSSGVEGRDVVALIRKAIQRRGDFDIDIVAVVNDTVGTMMTCGYDDHNCEIGLIVGTGSNACYMEEMRHIDMVEGDEGRMCINMEWGAFGDDGSLNDIRTEFDQEIDMGSLNPGKQLFEKMISGMYMGELVRLILVKMAKEELLFGGKLSPELLNTGRFETKDISDIEGEKDGIRKAREVLMRLGLDPTQEDCVATHRICQIVSTRSASLCAATLAAVLQRIKENKGEERLRSTIGVDGSVYKKHPHFAKRLHKTVRRLVPGCDVRFLRSEDGSGKGAAMVTAVAYRLADQHRARQKTLEHLQLSHDQLLEVKRRMKVEMERGLSKETHASAPVKMLPTYVCATPDGTEKGDFLALDLGGTNFRVLLVRVRNGKWGGVEMHNKIYAIPQEVMHGTGDELFDHIVQCIADFLEYMGMKGVSLPLGFTFSFPCQQNSLDESILLKWTKGFKASGCEGEDVVTLLKEAIHRREEFDLDVVAVVNDTVGTMMTCGFEDPHCEVGLIVGTGSNACYMEEMRNVELVEGEEGRMCVNMEWGAFGDNGCLDDFRTEFDVAVDELSLNPGKQRFEKMISGMYLGEIVRNILIDFTKRGLLFRGRISERLKTRGIFETKFLSQIESDCLALLQVRAILQHLGLESTCDDSIIVKEVCTVVARRAAQLCGAGMAAVVDRIRENRGLDALKVTVGVDGTLYKLHPHFAKVMHETVKDLAPKCDVSFLQSEDGSGKGAALITAVACRIREAGQR DIP protein P52789 original sequence version: 131 F17CE1938CF13880 src_mouse Neuronal proto-oncogene tyrosine-protein kinase Src Src pp60c-src Proto-oncogene c-Src protein protein mouse Mus musculus Mouse MGSNKSKPKDASQRRRSLEPSENVHGAGGAFPASQTPSKPASADGHRGPSAAFVPPAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMNKGSLLDFLKGETGKYLRLPQLVDMSAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL DIP protein P05480 original sequence version: 163 DIP protein P05480 original sequence version: 99 DIP protein P05480 original sequence version: 128 DIP protein P05480 original sequence version: 136 DIP protein P05480 original sequence version: 176 22CB5C6BE7A061C0 2-deoxy-d-glucose 2-deoxy-D-glucose 2-Deoxy-D-arabino-hexose 2-Deoxy-D-glucose 2-Deoxy-D-mannose D-2dGlc D-arabino-2-Deoxyhexose Deoxyglucose 2-deoxy-D-arabino-hexose small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) hxk1_human Hexokinase-1 Hexokinase type I Brain form hexokinase HK1 Hexokinase-A protein protein human Homo sapiens Human MIAAQLLAYYFTELKDDQVKKIDKYLYAMRLSDETLIDIMTRFRKEMKNGLSRDFNPTATVKMLPTFVRSIPDGSEKGDFIALDLGGSSFRILRVQVNHEKNQNVHMESEVYDTPENIVHGSGSQLFDHVAECLGDFMEKRKIKDKKLPVGFTFSFPCQQSKIDEAILITWTKRFKASGVEGADVVKLLNKAIKKRGDYDANIVAVVNDTVGTMMTCGYDDQHCEVGLIIGTGTNACYMEELRHIDLVEGDEGRMCINTEWGAFGDDGSLEDIRTEFDREIDRGSLNPGKQLFEKMVSGMYLGELVRLILVKMAKEGLLFEGRITPELLTRGKFNTSDVSAIEKNKEGLHNAKEILTRLGVEPSDDDCVSVQHVCTIVSFRSANLVAATLGAILNRLRDNKGTPRLRTTVGVDGSLYKTHPQYSRRFHKTLRRLVPDSDVRFLLSESGSGKGAAMVTAVAYRLAEQHRQIEETLAHFHLTKDMLLEVKKRMRAEMELGLRKQTHNNAVVKMLPSFVRRTPDGTENGDFLALDLGGTNFRVLLVKIRSGKKRTVEMHNKIYAIPIEIMQGTGEELFDHIVSCISDFLDYMGIKGPRMPLGFTFSFPCQQTSLDAGILITWTKGFKATDCVGHDVVTLLRDAIKRREEFDLDVVAVVNDTVGTMMTCAYEEPTCEVGLIVGTGSNACYMEEMKNVEMVEGDQGQMCINMEWGAFGDNGCLDDIRTHYDRLVDEYSLNAGKQRYEKMISGMYLGEIVRNILIDFTKKGFLFRGQISETLKTRGIFETKFLSQIESDRLALLQVRAILQQLGLNSTCDDSILVKTVCGVVSRRAAQLCGAGMAAVVDKIRENRGLDRLNVTVGVDGTLYKLHPHFSRIMHQTVKELSPKCNVSFLLSEDGSGKGAALITAVGVRLRTEASS DIP protein P19367 original sequence version: 188 F29A6837531C0594 src_human Proto-oncogene tyrosine-protein kinase Src pp60c-src SRC SRC1 Proto-oncogene c-Src protein protein human Homo sapiens Human MGSNKSKPKDASQRRRSLEPAENVHGAGGGAFPASQTPSKPASADGHRGPSAAFAPAAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKGETGKYLRLPQLVDMAAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL DIP protein P12931 original sequence version: 188 DIP protein P12931 original sequence version: 189 DIP protein P12931 original sequence version: 190 DIP protein P12931 original sequence version: 191 DIP protein P12931 original sequence version: 115 DIP protein P12931 original sequence version: 116 DIP protein P12931 original sequence version: 164 DIP protein P12931 original sequence version: 142 DIP protein P12931 original sequence version: 143 DIP protein P12931 original sequence version: 150 DIP protein P12931 original sequence version: 160 DIP protein P12931 original sequence version: 184 DIP protein P12931 original sequence version: 155 DIP protein P12931-1 original sequence version: 155 DIP protein P12931 original sequence version: 202 C1908084683E5DE8 hk1-src-1 2 11 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 9 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Among ten human glycolytic enzymes co-expressed individually with HA-c-Src, HK1 was exclusively precipitated by HA-c-Src (Fig. 1a). Fig. 1A, 1C src-hk1-1 2 11 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 region nphosres N-phosphorylated residue NPhosRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined observed-ptm observed-ptm observed ptm observed-ptm P19367:p.Tyr27Phe mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 19:29:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr27phe' Tue Jun 30 15:54:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr27Phe' P19367:p.Tyr417Phe mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 17:45:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr417phe' Tue Jun 30 17:42:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr417Phe' P19367:p.Tyr732Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 19:51:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr732phe' Tue Jun 30 17:48:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr732Phe' ser-732 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain Y F prerequisite-ptm prerequisite-ptm prerequisite ptm required ptm prerequisite-ptm required-ptm 9 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined sh2 region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain P05480:p.Tyr521Phe mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 14:30:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr527phe' Tue Jun 30 17:37:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr527Phe' Mon Mar 01 11:33:26 GMT 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'P05480:p.Tyr527Phe' physical association physical association Fig. 1B, 2A, 2B, 2C, 2G, 2I, Supp. 1A, 1C, 1D, 3H Kinase-dead SRC failed to phosphorylate HK1 and phosphorylation was inhibited by c-Src inhibitors PP2 and SU6656, but enhanced by stimulating cells with PDGF. CHEBI:78331 -PP2 CHEBI:91462 -SU6656 hk1-src-2 1 11 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait physical association physical association Fig. 1D Endogenous c-Src in lysate of HCT116 cells was precipitated with anti-c-Src, followed by WB to detect c-Src and HK1. hk1-src-3 7 11 unspecified role unspecified role prey prey n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association In vitro GST-pull down assay also confirmed the direct interaction between His-HK1 and GST-c-Src, as indicated by coomassie brilliant blue staining (Fig. 1e, left panel) and western blot (Fig. 1e, right panel) Fig. 1E hk1-src-4 3 11 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 9 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization We also found strong co-localization between c-Src and HK1 in cytosol (Fig. 1f). Fig. 1F src-hk1-3 4 11 enzyme target enzyme target substrate substrate unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 region optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy western blot western blot Immuno blot Immuno blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined resulting-ptm resulting-ptm resulting ptm resulting-ptm P19367:p.Tyr732Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 17:41:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr732phe' Tue Jun 30 17:17:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr732Phe' tyr732 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy western blot western blot Immuno blot Immuno blot mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) 9 enzyme enzyme unspecified role unspecified role n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 2D, 2H, 2J HK1 is phosphorylated by c-Src at Tyr732. P05480 (GO:0004672) up-regulates activity P19367 (GO:0004396) P05480 (GO:0004672) up-regulates activity P19367 (GO:0005536) hk1-2_deoxy_d_glucose-1 5 10 unspecified role unspecified role prey prey region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 4B hk1-hk1-1 2 11 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region nphosres N-phosphorylated residue NPhosRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined prerequisite-ptm prerequisite-ptm prerequisite ptm required ptm prerequisite-ptm required-ptm 11 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 region nphosres N-phosphorylated residue NPhosRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined prerequisite-ptm prerequisite-ptm prerequisite ptm required ptm prerequisite-ptm required-ptm physical association physical association Fig. 4C-E Src hk1-hk1-2 2 9 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region nphosres N-phosphorylated residue NPhosRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined prerequisite-ptm prerequisite-ptm prerequisite ptm required ptm prerequisite-ptm required-ptm 11 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 region nphosres N-phosphorylated residue NPhosRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined prerequisite-ptm prerequisite-ptm prerequisite ptm required ptm prerequisite-ptm required-ptm association association We next carried out two-step co-IP and found that c-Src could associate with HK1 dimer to form a ternary complex (Fig. 4f). Fig. 4F Two-step co-IP was performed, first IPs were carried out with anti-Flag, followed by anti-Myc. hk1-src-5 6 11 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component physical association physical association Fig.4G src-hk1-2 2 11 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 9 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Supp Fig. 1B hk2-src-1 2 8 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 9 unspecified role unspecified role prey prey P05480:p.Tyr521Phe mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 20:21:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr527phe' Tue Jun 30 18:59:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr527Phe' Mon Mar 01 10:52:22 GMT 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'P05480:p.Tyr527Phe' region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Supp Fig. 1G, 1I, 1J Kinase-dead SRC failed to phosphorylate HK2 and phosphorylation was inhibited by c-Src inhibitor PP2 src-hk2-1 7 8 unspecified role unspecified role prey prey n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Supp Fig. 1H src-hk2-2 4 8 enzyme target enzyme target substrate substrate unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 region optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy western blot western blot Immuno blot Immuno blot undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined resulting-ptm resulting-ptm resulting ptm resulting-ptm P52789:p.Tyr686Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 17:16:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr686phe' Tue Jun 30 18:29:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr686Phe' tyr732 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy western blot western blot Immuno blot Immuno blot mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) 9 enzyme enzyme unspecified role unspecified role n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Supp Fig 2I, 2K hk1-src-6 3 11 unspecified role unspecified role neutral component neutral component region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 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 Supp. Fig. 1E hk2-src-3 2 8 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 9 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Supp Fig. 1F