mbinfo MBInfo http://www.mechanobio.info/ http://www.mechanobio.info/ lim-2003-3 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. 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 J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation lim-2003-1 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. 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 J. Biol. Chem. (0021-9258) 2003 Only protein-protein interactions imex curation lim-2003-2 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. 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 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 kpca_human Protein kinase C alpha type PRKCA PKCA PRKACA protein protein human Homo sapiens Human MADVFPGNDSTASQDVANRFARKGALRQKNVHEVKDHKFIARFFKQPTFCSHCTDFIWGFGKQGFQCQVCCFVVHKRCHEFVTFSCPGADKGPDTDDPRSKHKFKIHTYGSPTFCDHCGSLLYGLIHQGMKCDTCDMNVHKQCVINVPSLCGMDHTEKRGRIYLKAEVADEKLHVTVRDAKNLIPMDPNGLSDPYVKLKLIPDPKNESKQKTKTIRSTLNPQWNESFTFKLKPSDKDRRLSVEIWDWDRTTRNDFMGSLSFGVSELMKMPASGWYKLLNQEEGEYYNVPIPEGDEEGNMELRQKFEKAKLGPAGNKVISPSEDRKQPSNNLDRVKLTDFNFLMVLGKGSFGKVMLADRKGTEELYAIKILKKDVVIQDDDVECTMVEKRVLALLDKPPFLTQLHSCFQTVDRLYFVMEYVNGGDLMYHIQQVGKFKEPQAVFYAAEISIGLFFLHKRGIIYRDLKLDNVMLDSEGHIKIADFGMCKEHMMDGVTTRTFCGTPDYIAPEIIAYQPYGKSVDWWAYGVLLYEMLAGQPPFDGEDEDELFQSIMEHNVSYPKSLSKEAVSVCKGLMTKHPAKRLGCGPEGERDVREHAFFRRIDWEKLENREIQPPFKPKVCGKGAENFDKFFTRGQPVLTPPDQLVIANIDQSDFEGFSYVNPQFVHPILQSAV sdc4_rat Syndecan-4 Sdc4 Synd4 Ryudocan core protein protein protein rat Rattus norvegicus (Rat) Rat MAPVCLFAPLLLLLLGGFPVAPGESIRETEVIDPQDLLEGRYFSGALPDDEDAGGLEQDSDFELSGSGDLDDTEEPRTFPEVISPLVPLDNHIPENAQPGIRVPSEPKELEENEVIPKRVPSDVGDDDVSNKVSMSSTSQGSNIFERTEVLAALIVGGVVGILFAVFLILLLVYRMKKKDEGSYDLGKKPIYKKAPTNEFYA sdc4_human Syndecan-4 SDC4 Amphiglycan Ryudocan core protein protein protein human Homo sapiens Human MAPARLFALLLFFVGGVAESIRETEVIDPQDLLEGRYFSGALPDDEDVVGPGQESDDFELSGSGDLDDLEDSMIGPEVVHPLVPLDNHIPERAGSGSQVPTEPKKLEENEVIPKRISPVEESEDVSNKVSMSSTVQGSNIFERTEVLAALIVGGIVGILFAVFLILLLMYRMKKKDEGSYDLGKKPIYKKAPTNEFYA sdc4-prkca-5 1287203 n/a 1287206 unspecified role unspecified role prey prey n/a 1287207 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 false catalytic subdomain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1287208 unspecified role unspecified role bait bait association association false false false 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-8 1287203 n/a 1287208 unspecified role unspecified role bait bait n/a 1287206 unspecified role unspecified role prey prey association association false false false 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 1287203 n/a 1287208 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 false n/a 1287206 unspecified role unspecified role prey prey physical association physical association false false false Fig 6d. sdc4-prkca-1 1287204 n/a 1287208 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 false P34901:p.Tyr192Phe mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n/a 1287207 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 false 1287228 1287225 physical association physical association false false false Fig 1c, 1d, 2a, 2b and 6d. sdc4-sdc4 1287204 n/a 1287208 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 false P34901:p.Ser183Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n/a 1287208 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 false 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 false 1287235 1287231 physical association physical association false false false Fig 4a and 4b. sdc4-prkca-2 1287205 n/a 1287207 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 false 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 false 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. n/a 1287209 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 false chemical synthesis Chemical synthesis (Chemical synthesis) 1287241 1287238 direct interaction direct interaction false false false Fig 3a. His6 and Xpress-tagged PKC-alpha detected by anti-Xpress antibodies. sdc4-prkca-3 1287205 n/a 1287207 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 false 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 false 9bact Bacteria n/a 1287209 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 false chemical synthesis Chemical synthesis (Chemical synthesis) 1287247 1287245 direct interaction direct interaction false false false Fig 3b.