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/ Structural basis for activation of the receptor tyrosine kinase KIT by stem cell factor. Structural basis for activation of the receptor tyrosine kinase KIT by stem cell factor. Cell (0092-8674) 2007 imex curation Yuzawa S., Opatowsky Y., Zhang Z., Mandiyan V., Lax I., Schlessinger J. 3T3 cells expressing KIT were treated with 125I-SCF in the presence of increasing concentrations of native SCF. joseph.schlessinger@yale.edu 07-Sept-2007: Contacted by JYOTI. davet Only protein-protein interactions 2 in vitro In vitro competition binding competition binding predetermined predetermined participant predetermined Yuzawa S., Opatowsky Y., Zhang Z., Mandiyan V., Lax I., Schlessinger J. 3T3 cells expressing KIT were treated with 125I-SCF in the presence of increasing concentrations of native SCF. joseph.schlessinger@yale.edu Accepted 2007-SEP-8 by DAVET. 07-Sept-2007: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 2 imex curation Structural basis for activation of the receptor tyrosine kinase KIT by stem cell factor. Structural basis for activation of the receptor tyrosine kinase KIT by stem cell factor. Cell (0092-8674) 2007 imex curation Yuzawa S., Opatowsky Y., Zhang Z., Mandiyan V., Lax I., Schlessinger J. 3T3 cells expressing KIT were treated with 125I-SCF in the presence of increasing concentrations of native SCF. joseph.schlessinger@yale.edu 07-Sept-2007: Contacted by JYOTI. davet Only protein-protein interactions 2 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Yuzawa S., Opatowsky Y., Zhang Z., Mandiyan V., Lax I., Schlessinger J. 3T3 cells expressing KIT were treated with 125I-SCF in the presence of increasing concentrations of native SCF. joseph.schlessinger@yale.edu Accepted 2007-SEP-8 by DAVET. 07-Sept-2007: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 2 imex curation scf_human Kit ligand c-Kit ligand Stem cell factor Mast cell growth factor KITLG MGF SCF protein protein human Homo sapiens Human MKKTQTWILTCIYLQLLLFNPLVKTEGICRNRVTNNVKDVTKLVANLPKDYMITLKYVPGMDVLPSHCWISEMVVQLSDSLTDLLDKFSNISEGLSNYSIIDKLVNIVDDLVECVKENSSKDLKKSFKSPEPRLFTPEEFFRIFNRSIDAFKDFVVASETSDCVVSSTLSPEKDSRVSVTKPFMLPPVAASSLRNDSSSSNRKAKNPPGDSSLHWAAMALPALFSLIIGFAFGALYWKKRQPSLTRAVENIQINEEDNEISMLQEKEREFQEV 19FD362CB59C6607 kit_human Mast/stem cell growth factor receptor Kit Tyrosine-protein kinase Kit KIT Proto-oncogene c-Kit SCFR Piebald trait protein p145 c-kit v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog protein protein human Homo sapiens Human MRGARGAWDFLCVLLLLLRVQTGSSQPSVSPGEPSPPSIHPGKSDLIVRVGDEIRLLCTDPGFVKWTFEILDETNENKQNEWITEKAEATNTGKYTCTNKHGLSNSIYVFVRDPAKLFLVDRSLYGKEDNDTLVRCPLTDPEVTNYSLKGCQGKPLPKDLRFIPDPKAGIMIKSVKRAYHRLCLHCSVDQEGKSVLSEKFILKVRPAFKAVPVVSVSKASYLLREGEEFTVTCTIKDVSSSVYSTWKRENSQTKLQEKYNSWHHGDFNYERQATLTISSARVNDSGVFMCYANNTFGSANVTTTLEVVDKGFINIFPMINTTVFVNDGENVDLIVEYEAFPKPEHQQWIYMNRTFTDKWEDYPKSENESNIRYVSELHLTRLKGTEGGTYTFLVSNSDVNAAIAFNVYVNTKPEILTYDRLVNGMLQCVAAGFPEPTIDWYFCPGTEQRCSASVLPVDVQTLNSSGPPFGKLVVQSSIDSSAFKHNGTVECKAYNDVGKTSAYFNFAFKGNNKEQIHPHTLFTPLLIGFVIVAGMMCIIVMILTYKYLQKPMYEVQWKVVEEINGNNYVYIDPTQLPYDHKWEFPRNRLSFGKTLGAGAFGKVVEATAYGLIKSDAAMTVAVKMLKPSAHLTEREALMSELKVLSYLGNHMNIVNLLGACTIGGPTLVITEYCCYGDLLNFLRRKRDSFICSKQEDHAEAALYKNLLHSKESSCSDSTNEYMDMKPGVSYVVPTKADKRRSVRIGSYIERDVTPAIMEDDELALDLEDLLSFSYQVAKGMAFLASKNCIHRDLAARNILLTHGRITKICDFGLARDIKNDSNYVVKGNARLPVKWMAPESIFNCVYTFESDVWSYGIFLWELFSLGSSPYPGMPVDSKFYKMIKEGFRMLSPEHAPAEMYDIMKTCWDADPLKRPTFKQIVQLIEKQISESTNHIYSNLANCSPNRQKPVVDHSVRINSVGSTASSSQPLLVHDDV DIP protein P10721 original sequence version: 149 81B0CD76817F3454 scf-kit-1 2 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component 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 ectodomain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 10 7 direct interaction direct interaction KIT was partially deglycosylated prior to crystallization The observed SCF-KIT 2:2 complex in the crystal lattice is consistent with experiments demonstrating that KIT dimerization is driven by the dimeric SCF ligand ([Philo et al., 1996] and [Lemmon et al., 1997]). 3 3.5A 24.9% bound and 29.5% free scf-kit-2 1 4 unspecified role unspecified role prey prey extracellular region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain mouse-3t3 Mus musculus embryo cells 3t3 Mouse embryo fibroblast cells 3 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) physical association physical association The EC50 (ligand concentration that displaces 50% of 125I-SCF bound to c-KIT) of SCF toward wild type KIT = 1.1 nM. Mutations in KIT (R381A, E386A0) which prevent dimerization or kinase activity (K263A) do not alter this value. Displacement assay of cell-bound 125I-SCF using native SCF. 3T3 cells expressing WT (), R381A (down triangle, filled), R381A/E386A (), or a kinase-negative KIT (up triangle, filled) were treated with 125I-SCF in the presence of increasing concentrations of native SCF. Fig. 7B