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/ The postsynaptic density proteins Homer and Shank form a polymeric network structure. The postsynaptic density proteins Homer and Shank form a polymeric network structure. Cell (0092-8674) 2009 imex curation Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 10 in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column anti tag western anti tag western blot anti tag western Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu Accepted 2009-JUN-27 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 10 imex curation The postsynaptic density proteins Homer and Shank form a polymeric network structure. The postsynaptic density proteins Homer and Shank form a polymeric network structure. Cell (0092-8674) 2009 imex curation Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 10 in vitro In vitro light scattering light scattering predetermined predetermined participant predetermined Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu Accepted 2009-JUN-27 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 10 imex curation The postsynaptic density proteins Homer and Shank form a polymeric network structure. The postsynaptic density proteins Homer and Shank form a polymeric network structure. Cell (0092-8674) 2009 imex curation Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 10 in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu Accepted 2009-JUN-27 by JYOTI. 21-Dec-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 10 imex curation The postsynaptic density proteins Homer and Shank form a polymeric network structure. The postsynaptic density proteins Homer and Shank form a polymeric network structure. Cell (0092-8674) 2009 imex curation Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu 21-Dec-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 10 in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati weight by comassie molecular weight estimation by coomasie staining weight by comassie Hayashi MK., Tang C., Verpelli C., Narayanan R., Stearns MH., Xu RM., Li H., Sala C., Hayashi Y. Purified proteins were mixed and ultra-centrifuged after overnight incubation. hayashim@mit.edu Accepted 2009-JUN-27 by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 10 imex curation shan1_rat SH3 and multiple ankyrin repeat domains protein 1 Shank1 GKAP/SAPAP-interacting protein Synamon Somatostatin receptor-interacting protein SPANK-1 protein protein rat Rattus norvegicus (Rat) Rat MTHSPATSEDEERHSASECPEGGSESDSSPDGPGRGPQGTRGRGSGAPGNLASTRGLQGRSMSVPDDAHFSMMVFRIGIPDLHQTKCLRFNPDATIWTAKQQVLCALSESLQDVLNYGLFQPATSGRDANFLEEERLLREYPQSFEKGVPYLEFRYKTRVYKQTNLDEKQLAKLHTKTGLKKFLEYVQLGTSDKVARLLDKGLDPNYHDSDSGETPLTLAAQTEGSVEVIRTLCLGGAHIDFRARDGMTALHKAACARHCLALTALLDLGGSPNYKDRRGLTPLFHTAMVGGDPRCCELLLYNRAQLGIADENGWQEIHQACQRGHSQHLEHLLFYGAEPGAQNASGNTALHICALYNKETCARILLYRGANKDVKNNNGQTPFQVAVIAGNFELGELIRNHREQDVVPFQESPKYAARRRGPPGAGLTVPPALLRANSDTSMALPDWMVFSAPGASSSGTPGPTSGPQGQSQPSAPSTKLSSGTLRSASSPRGARARSPSRGRHPEDAKRQPRGRPSSSGTPRDGPAGGTGGSGGPGGSLGSRGRRRKLYSAVPGRSFMAVKSYQAQGEGEISLSKGEKIKVLSIGEGGFWEGQVKGRVGWFPSDCLEEVANRSQEGRQESRSDKAKRLFRHYTVGSYDSFDAPSLIDGIDSGSDYIIKEKTVLLQKKDSEGFGFVLRGAKAQTPIEEFTPTPAFPALQYLESVDEGGVAWRAGLRMGDFLIEVNGQNVVKVGHRQVVNMIRQGGNTLMVKVVMVTRHPDMDEAVHKKASQQAKRLPPPAISLRSKSMTSELEEMVSPWKKKIEYEQQPAAVPSMEKKRTVYQMALNKLDEILAAAQQTISASESPGPGGLASLGKHRPKGFFATESSFDPHHRSQPSYDRPSFLPPGPGLMLRQKSIGAAEDDRPYLAPPAMKFSRSLSVPGSEDIPPPPTTSPPEPPYSTPPAPSSSGRLTPSPRGGPFNPSSGGPLPASSPSSFDGPSPPDTRGGGREKSLYHSAALPPAHHHPPHHHHHHAPPPQPHHHHAHPPHPPEMETGGSPDDPPPRLALGPQPSLRGWRGGGPSPTSGAPSPSHHSSSGGSSGPTQAPALRYFQLPPRAASAAMYVPARSGRGRKGPLVKQTKVEGEPQKGSIPSASSPTSPALPRSEPPPAGPSEKNSIPIPTIIIKAPSTSSSGRSSQGSSTEAEPPTQPDGAGGGGSSPSPAPATSPVPPSPSPVPTPASPSGPATLDFTSQFGAALVGAARREGGWQNEARRRSTLFLSTDAGDEDGGDSGLGPGGPPGPRLRHSKSIDEGMFSAEPYLRLESGGSSGGYGAYAAGSRAYGGSGSSSAFTSFLPPRPLVHPLTGKALDPASPLGLALAARERALKESSEGGGTPQPPPRPPSPRYDAPPPTLHHHSPHSPHSPHARHEPVLRLWGDPARRELGYRAGLGSQEKALTASPPAARRSLLHRLPPTAPGVGPLLLQLGPEPPTPHPGVSKAWRTAAPEEPERLPLHVRFLENCQARPPPAGTRGSSTEDGPGVPPPSPRRVLPTSPTSPRGNEENGLPLLVLPPPAPSVDVDDGEFLFAEPLPPPLEFSNSFEKPESPLTPGPPHPLPDPPSPATPLPAAPPPAVAAAPPTLDSTASSLTSYDSEVATLTQGAPAAPGDPPAPGPPAPAAPAPPAPQPGPDPPPGTDSGIEEVDSRSSSDHPLETISSASTLSSLSAEGGGNTGGVAGGGAGVASGTELLDTYVAYLDGQAFGGSGTPGPPYPPQLMTPSKLRGRALGTSGNLRPGPSGGLRDPVTPTSPTVSVTGAGTDGLLALSACPGPSTAGVAGGPVAVEPEVPPVPLPAASSLPRKLLPWEEGPGPPPPPLPGPLSQPQASALATVKASIISELSSKLQQFGGSSTAGGALPWARGGSGGSTDSHHGGASYIPERTSSLQRQRLSEDSQTSLLSKPSSSIFQNWPKPPLPPLPTGSGVSSSTAAAPGATSPSASSASASTRHLQGVEFEMRPPLLRRAPSPSLLPASDHKVSPAPRPSSLPILPSGPIYPGLFDIRSSPTGGAGGSTDPFAPVFVPPHPGISGGLGGALSGASRSLSPTRLLSLPPDKPFGAKPLGFWTKFDVADWLEWLGLSEHRAQFLDHEIDGSHLPALTKEDYVDLGVTRVGHRMNIDRALKFFLER 3F478B5A7B18BA86 q9z214-2 Homer protein homolog 1 Homer1 Homer Vesl PSD-Zip45 VASP/Ena-related gene up-regulated during seizure and LTP 1 1b protein protein rat Rattus norvegicus (Rat) Rat MGEQPIFSTRAHVFQIDPNTKKNWVPTSKHAVTVSYFYDSTRNVYRIISLDGSKAIINSTITPNMTFTKTSQKFGQWADSRANTVYGLGFSSEHHLSKFAEKFQEFKEAARLAKEKSQEKMELTSTPSQESAGGDLQSPLTPESINGTDDERTPDVTQNSEPRAEPAQNALPFSHSSAISKHWEAELATLKGNNAKLTAALLESTANVKQWKQQLAAYQEEAERLHKRVTELECVSSQANAVHSHKTELSQTVQELEETLKVKEEEIERLKQEIDNARELQEQRDSLTQKLQEVEIRNKDLEGQLSELEQRLEKSQSEQDAFRSNLKTLLEILDGKIFELTELRDNLAKLLECS C8F4579BFE3E69BF q9nsc5-1 Homer protein homolog 3 3a HOMER3 protein protein human Homo sapiens Human MSTAREQPIFSTRAHVFQIDPATKRNWIPAGKHALTVSYFYDATRNVYRIISIGGAKAIINSTVTPNMTFTKTSQKFGQWADSRANTVYGLGFASEQHLTQFAEKFQEVKEAARLAREKSQDGGELTSPALGLASHQVPPSPLVSANGPGEEKLFRSQSADAPGPTERERLKKMLSEGSVGEVQWEAEFFALQDSNNKLAGALREANAAAAQWRQQLEAQRAEAERLRQRVAELEAQAASEVTPTGEKEGLGQGQSLEQLEALVQTKDQEIQTLKSQTGGPREALEAAEREETQQKVQDLETRNAELEHQLRAMERSLEEARAERERARAEVGRAAQLLDVSLFELSELREGLARLAEAAP 9FCC62319FF5165A q9z214-3 Homer protein homolog 1 Homer1 Homer Vesl PSD-Zip45 VASP/Ena-related gene up-regulated during seizure and LTP 1 1a Vesl protein protein rat Rattus norvegicus (Rat) Rat MGEQPIFSTRAHVFQIDPNTKKNWVPTSKHAVTVSYFYDSTRNVYRIISLDGSKAIINSTITPNMTFTKTSQKFGQWADSRANTVYGLGFSSEHHLSKFAEKFQEFKEAARLAKEKSQEKMELTSTPSQESAGGDLQSPLTPESINGTDDERTPDVTQNSEPRAEPAQNALPFSHRYTFNSAIMIK 9845DA6E2574BF83 gkap1_rat G kinase-anchoring protein 1 Gkap1 protein protein rat Rattus norvegicus (Rat) Rat MASAVLSSVLTTASRFALLQVDSGSGSDSEPGKGKGRNNGKSQTLGNKSTANEKKREKRRKKKEQQQSEANELRNLAFKKIPQKSSHSVCNVQHELSLPNPVQKESREENWQEWRQRDEQLTSEMFEADLEKALLLSKLEYEEHRQDYENAENASTQTKVINKKDKRKNHQGKDKPLTVSLKDFQCEDHISKKTEESNSSQPLSHDGGFFNRLEDDVHKILVREKRREQLTEHNGTDNCPAPEHNQEVGLKDGRIERLKLELERKDAEIQKLKAVVTQWEAKYKEVKARNGQLLKMLQEGEMKDKAEILLQVDESQSIKNELTVQVSSLHAALEQERSKVKVLQAELAKYQGGRKGKRNCEPDQCR 435F1B1618361129 homer1-shank1-1 4 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component Q9Z214-2:p.Trp24Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W A Wed May 02 15:34:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp24ala' Tue Jun 30 17:45:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp24Ala' ecoli Escherichia coli (strain K12) physical association physical association In contrast, when they were mixed at a 1:1 molar ratio, approximately 50% of Shank1CΔPEST and Homer1b were precipitated, suggesting the formation of a high molecular weight complex. Figs 1B, 1C, 1E Mutation of a proline in the Homer Binding site of Shak1 prevented binding to wild-type Shak1 but this could not be mapped in this construct. homer1-shank1-2 2 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction On dynamic light scattering, a sensitive method for detecting large complex, Homer1b or Shank1CΔPEST alone had a single scattering peak corresponding to the hydrodynamic radii of 10.3 and 4.3 nm, respectively (Figure 1D). In contrast, the mixture of Homer1b and Shank1CΔPEST showed two peaks Figs 1D homer1-shank1-3 3 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction Electron microscopy images of Homer, Shank, and Homer-Shank complex. Figs 1I The Homer and Shank mixture showed a mesh-like structure in electron microscopic images. The mesh appears to contain hubs, possibly representing Shank oligomerized via its SAM domain and linked by the filamentous Homer. homer1-shank1-4 4 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-2339073]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 451-451, sequence length 366 [EBI-2339073]451-666 ecoli Escherichia coli (strain K12) physical association physical association When purified C-terminal fragment of GKAP (451�666) was incubated with Homer and Shank, the GKAP fragment coprecipitated with the Homer-Shank complex without affecting the amount of Homer or Shank in the precipitates (Figures 2A and 2B). Figs 2A, 2B homer1-shank1-5 4 8 competitor competitor competitor neutral component neutral component ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component Q9Z214-2:p.Trp24Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W A Wed May 02 19:18:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp24ala' Tue Jun 30 19:03:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp24Ala' ecoli Escherichia coli (strain K12) physical association physical association Dose-dependent inhibition of the high-order complex formation between Homer and Shank by Homer1a Figs 2C, 2D A competitive interaction between Homer1a and Homer1b on Shank prevents the complex formation. homer3-shank1-1 4 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction By adding the purified CaMKIIα activated with Ca2+ and calmodulin to the mixture of Homer and Shank (Figures 2E and 2F), the complex formation between Homer3a and Shank1CΔPEST was inhibited, Figs 2E, 2F Interaction is inhibited by phosphorylation of Homer3 homer1-1 4 6 unspecified role unspecified role neutral component neutral component Q9Z214-2:p.[Ile332Arg;Ile337Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain I R certain certain sequence position certain certain certain sequence position certain I E Wed May 02 17:40:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile332arg,ile337glu' Tue Jun 30 16:14:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile332Arg;Ile337Glu]' region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Representative scans of sedimentation equilibrium analysis of Homer mutants done at centrifuge speed of 12,000 rpm Fig. 4C, 4F homer1-2 1 6 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component Q9Z214-2:p.[Ile332Arg;Ile337Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain I R certain certain sequence position certain certain certain sequence position certain I E Wed May 02 19:14:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile332arg,ile337glu' Tue Jun 30 17:25:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile332Arg;Ile337Glu]' region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction This shows that the dimeric mutant interacts with wild-type Homer to make heteromeric dimers, although some of the heteromers still form tetramers. Fig. 4C, 4F homer1-3 1 6 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction Individually expressed HA-Homer1b and myc-Homer1b I332R/I337E were eluted as a single peak at the expected position of tetramer and dimer, respectively. Fig. 4D homer1-4 2 6 unspecified role unspecified role neutral component neutral component Q9Z214-2:p.[Ile332Arg;Ile337Glu] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain I R certain certain sequence position certain certain certain sequence position certain I E Wed May 02 18:49:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile332arg,ile337glu' Tue Jun 30 16:05:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile332Arg;Ile337Glu]' region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Individually expressed HA-Homer1b and myc-Homer1b I332R/I337E were eluted as a single peak at the expected position of tetramer and dimer, respectively. Fig. 4D