UniProt UniProt http://www.uniprot.org Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Oncogene (0950-9232) 2011 imex curation Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu 20-Jun-2012: Contacted by JYOTI. 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 Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu Accepted 2012-JUN-06 by ORCHARD Oncogene (0950-9232) 2011 Only protein-protein interactions imex curation Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Oncogene (0950-9232) 2011 imex curation Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu Accepted 2012-JUN-06 by ORCHARD 20-Jun-2012: Contacted by JYOTI. Oncogene (0950-9232) 2011 Only protein-protein interactions imex curation Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Oncogene (0950-9232) 2011 imex curation Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions human-du_145 Homo sapiens DU-145 prostate cancer cell line du_145 DU-145 Human prostate carcinoma established from the tumor tissue removed from the metastatic central nervous system lesion of a 69-year-old man with prostate carcinoma in 1975 confirmed as human with IEF of AST, MDH, NP anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu Accepted 2012-JUN-06 by ORCHARD 20-Jun-2012: Contacted by JYOTI. Oncogene (0950-9232) 2011 Only protein-protein interactions imex curation Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Oncogene (0950-9232) 2011 imex curation Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions human-cwr22rv1 Homo sapiens cwr22rv1 Androgen-responsive human prostate carcinoma cell line CWR22Rv1 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu Accepted 2012-JUN-06 by ORCHARD 20-Jun-2012: Contacted by JYOTI. Oncogene (0950-9232) 2011 Only protein-protein interactions imex curation Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Male germ cell-associated kinase is overexpressed in prostate cancer cells and causes mitotic defects via deregulation of APC/C(CDH1). Oncogene (0950-9232) 2011 imex curation Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay autoradiography autoradiography Wang L.Y., Kung H.J. Cancer - Interactions investigated in the context of cancer hkung@ucdavis.edu Accepted 2012-JUN-06 by ORCHARD 20-Jun-2012: Contacted by JYOTI. Oncogene (0950-9232) 2011 Only protein-protein interactions imex curation fzr1_human Fizzy-related protein homolog FYR FZR KIAA1242 FZR1 CDH1 Cdh1/Hct1 homolog CDC20-like protein 1 protein protein human Homo sapiens Human MDQDYERRLLRQIVIQNENTMPRVTEMRRTLTPASSPVSSPSKHGDRFIPSRAGANWSVNFHRINENEKSPSQNRKAKDATSDNGKDGLAYSALLKNELLGAGIEKVQDPQTEDRRLQPSTPEKKGLFTYSLSTKRSSPDDGNDVSPYSLSPVSNKSQKLLRSPRKPTRKISKIPFKVLDAPELQDDFYLNLVDWSSLNVLSVGLGTCVYLWSACTSQVTRLCDLSVEGDSVTSVGWSERGNLVAVGTHKGFVQIWDAAAGKKLSMLEGHTARVGALAWNAEQLSSGSRDRMILQRDIRTPPLQSERRLQGHRQEVCGLKWSTDHQLLASGGNDNKLLVWNHSSLSPVQQYTEHLAAVKAIAWSPHQHGLLASGGGTADRCIRFWNTLTGQPLQCIDTGSQVCNLAWSKHANELVSTHGYSQNQILVWKYPSLTQVAKLTGHSYRVLYLAMSPDGEAIVTGAGDETLRFWNVFSKTRSTKVKWESVSVLNLFTRIR DIP protein Q9UM11 original sequence version: 100 DIP protein Q9UM11 original sequence version: 131 DIP protein Q9UM11 original sequence version: 136 DIP protein Q9UM11 original sequence version: 145 F6C8FDAF81D8103A cdk20_human Cyclin-dependent kinase 20 CDK20 CCRK CDCH CDK-activating kinase p42 Cell cycle-related kinase Cell division protein kinase 20 Cyclin-dependent protein kinase H Cyclin-kinase-activating kinase p42 protein protein human Homo sapiens Human MDQYCILGRIGEGAHGIVFKAKHVETGEIVALKKVALRRLEDGFPNQALREIKALQEMEDNQYVVQLKAVFPHGGGFVLAFEFMLSDLAEVVRHAQRPLAQAQVKSYLQMLLKGVAFCHANNIVHRDLKPANLLISASGQLKIADFGLARVFSPDGSRLYTHQVATRWYRAPELLYGARQYDQGVDLWSVGCIMGELLNGSPLFPGKNDIEQLCYVLRILGTPNPQVWPELTELPDYNKISFKEQVPMPLEEVLPDVSPQALDLLGQFLLYPPHQRIAASKALLHQYFFTAPLPAHPSELPIPQRLGGPAPKAHPGPPHIHDFHVDRPLEESLLNPELIRPFILEG EB6A53E97301D5AE mak_human Serine/threonine-protein kinase MAK MAK Male germ cell-associated kinase protein protein human Homo sapiens Human MNRYTTMRQLGDGTYGSVLMGKSNESGELVAIKRMKRKFYSWDECMNLREVKSLKKLNHANVIKLKEVIRENDHLYFIFEYMKENLYQLMKDRNKLFPESVIRNIMYQILQGLAFIHKHGFFHRDMKPENLLCMGPELVKIADFGLARELRSQPPYTDYVSTRWYRAPEVLLRSSVYSSPIDVWAVGSIMAELYMLRPLFPGTSEVDEIFKICQVLGTPKKSDWPEGYQLASSMNFRFPQCVPINLKTLIPNASNEAIQLMTEMLNWDPKKRPTASQALKHPYFQVGQVLGPSSNHLESKQSLNKQLQPLESKPSLVEVEPKPLPDIIDQVVGQPQPKTSQQPLQPIQPPQNLSVQQPPKQQSQEKPPQTLFPSIVKNMPTKPNGTLSHKSGRRRWGQTIFKSGDSWEELEDYDFGASHSKKPSMGVFKEKRKKDSPFRLPEPVPSGSNHSTGENKSLPAVTSLKSDSELSTAPTSKQYYLKQSRYLPGVNPKKVSLIASGKEINPHTWSNQLFPKSLGPVGAELAFKRSNAGNLGSYATYNQSGYIPSFLKKEVQSAGQRIHLAPLNATASEYTWNTKTGRGQFSGRTYNPTAKNLNIVNRAQPIPSVHGRTDWVAKYGGHR 67F540266F370285 mak-cdk20 1 8 unspecified role unspecified role bait bait flag-mak flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 7 unspecified role unspecified role prey prey ha-cdk20 ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 3D mak-fzr1 1 6 unspecified role unspecified role prey prey ha-fzr1 ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag anti tag western anti tag western blot anti tag western n-term range n-terminal range n-term range n-term range n-terminal range n-term range 8 unspecified role unspecified role bait bait v5-mak v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag anti tag western anti tag western blot anti tag western n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 4A mak-fzr1-2 3 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Figure 4B mak-fzr1-3 4 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association Fig 4B mak-fzr1-4 2 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association Fig 4B fzr1-mak 5 8 enzyme enzyme unspecified role unspecified role v5-mak v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope 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. 6 enzyme target enzyme target substrate substrate unspecified role unspecified role gst-fzr1 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) phosphorylation phosphorylation reaction phosphorylation Figure 5a fzr1-mak-1 5 6 enzyme target enzyme target substrate substrate unspecified role unspecified role gst-fzr1 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) 8 enzyme enzyme unspecified role unspecified role v5-mak v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. phosphorylation phosphorylation reaction phosphorylation Figure 5b MAK phosphorylates CDH1 during G2 to M phase. fzr1-mak-2 5 8 enzyme enzyme unspecified role unspecified role gst-mak 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) 6 enzyme target enzyme target substrate substrate unspecified role unspecified role gst-fzr1 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) phosphorylation phosphorylation reaction phosphorylation Figure 5c Mutant CDH1(4A) lacks the four major phosphoacceptor sites of CDK (S40A, T121A, S151A, S163A) and mutant CDH1(9A) lacks all nine acceptor sites (T32, S36, S40, S70, T121, S138, S146, S151, S163). Phosphorylation of CDH1(4A) by MAK was significantly diminished, and only marginal phosphorylation of CDH1(9A) could be detected