mbinfo MBInfo http://www.mechanobio.info/ Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. J. Biol. Chem. (0021-9258) 2003 imex curation Lim S.T., Longley R.L., Couchman J.R., Woods A. The source of PKC-alpha specified by the author and the source of syndecan-4 inferred from the amino acid positions and the use of rat syndecan-4 template in site-directed mutagenesis. anwoods@uab.edu 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Lim S.T., Longley R.L., Couchman J.R., Woods A. Agarose pull down was used. The source of PKC-alpha specified by the author and the source of syndecan-4 inferred from the amino acid positions and the use of rat syndecan-4 template in site-directed mutagenesis. anwoods@uab.edu Accepted 2012-APR-30 by ORCHARD 20-Jun-2012: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. J. Biol. Chem. (0021-9258) 2003 imex curation Lim S.T., Longley R.L., Couchman J.R., Woods A. The source of PKC-alpha specified by the author and the source of syndecan-4 inferred from the amino acid positions and the use of rat syndecan-4 template in site-directed mutagenesis. anwoods@uab.edu 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions yeast Saccharomyces cerevisiae 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 predetermined predetermined participant predetermined Lim S.T., Longley R.L., Couchman J.R., Woods A. The source of PKC-alpha specified by the author and the source of syndecan-4 inferred from the amino acid positions and the use of rat syndecan-4 template in site-directed mutagenesis. anwoods@uab.edu Accepted 2012-APR-30 by ORCHARD J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. Direct binding of syndecan-4 cytoplasmic domain to the catalytic domain of protein kinase C alpha (PKC alpha) increases focal adhesion localization of PKC alpha. J. Biol. Chem. (0021-9258) 2003 imex curation Lim S.T., Longley R.L., Couchman J.R., Woods A. The source of PKC-alpha specified by the author and the source of syndecan-4 inferred from the amino acid positions and the use of rat syndecan-4 template in site-directed mutagenesis. anwoods@uab.edu 20-Jun-2012: Contacted by JYOTI. Only protein-protein interactions rat-ref52 Rattus norvegicus embryonic fibroblast cells ref52 Rattus norvegicus embryonic fibroblast cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lim S.T., Longley R.L., Couchman J.R., Woods A. The source of PKC-alpha specified by the author and the source of syndecan-4 inferred from the amino acid positions and the use of rat syndecan-4 template in site-directed mutagenesis. anwoods@uab.edu Accepted 2012-APR-30 by ORCHARD 20-Jun-2012: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation kpca_rat Protein kinase C alpha type Prkca Pkca protein protein rat Rattus norvegicus (Rat) Rat MADVYPANDSTASQDVANRFARKGALRQKNVHEVKDHKFIARFFKQPTFCSHCTDFIWGFGKQGFQCQVCCFVVHKRCHEFVTFSCPGADKGPDTDDPRSKHKFKIHTYGSPTFCDHCGSLLYGLIHQGMKCDTCDMNVHKQCVINVPSLCGMDHTEKRGRIYLKAEVTDEKLHVTVRDAKNLIPMDPNGLSDPYVKLKLIPDPKNESKQKTKTIRSTLNPQWNESFTFKLKPSDKDRRLSVEIWDWDRTTRNDFMGSLSFGVSELMKMPASGWYKLLNQEEGEYYNVPIPEGDEEGNVELRQKFEKAKLGPAGNKVISPSEDRKQPSNNLDRVKLTDFNFLMVLGKGSFGKVMLADRKGTEELYAIKILKKDVVIQDDDVECTMVEKRVLALLDKPPFLTQLHSCFQTVDRLYFVMEYVNGGDLMYHIQQVGKFKEPQAVFYAAEISIGLFFLHKRGIIYRDLKLDNVMLDSEGHIKIADFGMCKEHMMDGVTTRTFCGTPDYIAPEIIAYQPYGKSVDWWAYGVLLYEMLAGQPPFDGEDEDELFQSIMEHNVSYPKSLSKEAVSICKGLMTKHPAKRLGCGPEGERDVREHAFFRRIDWEKLENREIQPPFKPKVCGKGAENFDKFFTRGQPVLTPPDQLVIANIDQSDFEGFSYVNPQFVHPILQSAV 94889E7339C17719 sdc4_human Syndecan-4 SDC4 Amphiglycan Ryudocan core protein protein protein human Homo sapiens Human MAPARLFALLLFFVGGVAESIRETEVIDPQDLLEGRYFSGALPDDEDVVGPGQESDDFELSGSGDLDDLEDSMIGPEVVHPLVPLDNHIPERAGSGSQVPTEPKKLEENEVIPKRISPVEESEDVSNKVSMSSTVQGSNIFERTEVLAALIVGGIVGILFAVFLILLLMYRMKKKDEGSYDLGKKPIYKKAPTNEFYA DIP protein P31431 original sequence version: 87 DIP protein P31431-1 original sequence version: 134 8229AA2733F77A10 sdc4_rat Syndecan-4 Sdc4 Synd4 Ryudocan core protein protein protein rat Rattus norvegicus (Rat) Rat MAPVCLFAPLLLLLLGGFPVAPGESIRETEVIDPQDLLEGRYFSGALPDDEDAGGLEQDSDFELSGSGDLDDTEEPRTFPEVISPLVPLDNHIPENAQPGIRVPSEPKELEENEVIPKRVPSDVGDDDVSNKVSMSSTSQGSNIFERTEVLAALIVGGVVGILFAVFLILLLVYRMKKKDEGSYDLGKKPIYKKAPTNEFYA 5D9C8B3BF4C0D58A kpca_human Protein kinase C alpha type PRKCA PKCA PRKACA protein protein human Homo sapiens Human MADVFPGNDSTASQDVANRFARKGALRQKNVHEVKDHKFIARFFKQPTFCSHCTDFIWGFGKQGFQCQVCCFVVHKRCHEFVTFSCPGADKGPDTDDPRSKHKFKIHTYGSPTFCDHCGSLLYGLIHQGMKCDTCDMNVHKQCVINVPSLCGMDHTEKRGRIYLKAEVADEKLHVTVRDAKNLIPMDPNGLSDPYVKLKLIPDPKNESKQKTKTIRSTLNPQWNESFTFKLKPSDKDRRLSVEIWDWDRTTRNDFMGSLSFGVSELMKMPASGWYKLLNQEEGEYYNVPIPEGDEEGNMELRQKFEKAKLGPAGNKVISPSEDRKQPSNNLDRVKLTDFNFLMVLGKGSFGKVMLADRKGTEELYAIKILKKDVVIQDDDVECTMVEKRVLALLDKPPFLTQLHSCFQTVDRLYFVMEYVNGGDLMYHIQQVGKFKEPQAVFYAAEISIGLFFLHKRGIIYRDLKLDNVMLDSEGHIKIADFGMCKEHMMDGVTTRTFCGTPDYIAPEIIAYQPYGKSVDWWAYGVLLYEMLAGQPPFDGEDEDELFQSIMEHNVSYPKSLSKEAVSVCKGLMTKHPAKRLGCGPEGERDVREHAFFRRIDWEKLENREIQPPFKPKVCGKGAENFDKFFTRGQPVLTPPDQLVIANIDQSDFEGFSYVNPQFVHPILQSAV 9EB157789A062349 sdc4-prkca-1 2 6 unspecified role unspecified role bait bait variable domain required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain P34901:p.Tyr192Phe mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 18:42:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr192phe' Tue Jun 30 17:52:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr192Phe' 7 unspecified role unspecified role prey prey catalytic subdomain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 13 10 physical association physical association Fig 1c, 1d, 2a, 2b and 6d. sdc4-prkca-2 1 7 unspecified role unspecified role prey prey catalytic domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 PKC-alpha was tagged with His6 and Xpress tags. drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. 5 unspecified role unspecified role bait bait variable domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 16 19 direct interaction direct interaction Fig 3a. His6 and Xpress-tagged PKC-alpha detected by anti-Xpress antibodies. sdc4-prkca-3 1 7 unspecified role unspecified role prey prey 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 catalytic subdomain required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 9bact Bacteria 5 unspecified role unspecified role bait bait variable domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 23 25 direct interaction direct interaction Fig 3b. sdc4-sdc4 2 6 unspecified role unspecified role putative self putative self variable domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain P34901:p.Ser183Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S D Wed May 02 19:26:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser183asp' Tue Jun 30 16:19:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser183Asp' 6 unspecified role unspecified role putative self putative self P34901:p.Ser183Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S D Wed May 02 14:38:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser183asp' Tue Jun 30 20:13:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser183Asp' variable domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 28 32 physical association physical association Fig 4a and 4b. sdc4-prkca-5 3 4 unspecified role unspecified role prey prey 7 competitor competitor competitor 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 catalytic subdomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait association association Fig 5a and 5b. PKC-alpha bound to syndecan-4 in the present of the competitor (GST-PKM5) was detected with anti-PKC-alpha. The actual displacement and binding of the competitor was detected by reprobing with anti-GST. sdc4-prkca-7 3 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role prey prey physical association physical association Fig 6d. sdc4-prkca-8 3 6 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey association association Fig 5a and 5b. PKC-alpha bound to syndecan-4 in the present of the competitor (GST-PKM5) was detected with anti-PKC-alpha. The actual displacement and binding of the competitor was detected by reprobing with anti-GST.