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/ http://www.ebi.ac.uk/ wang-2004d-2 Heterodimeric interaction and interfaces of S100A1 and S100P. yeasx Saccharomyces cerevisiae (Baker's yeast) 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 Wang G., Zhang S., Fernig DG., Spiller D., Martin-Fernandez M., Zhang H., Ding Y., Rao Z., Rudland PS., Barraclough R. Cancer - Interactions investigated in the context of cancer More than 2 E6 yeast cell transformants from a yeast two hybrid cDNA library, constructed from mRNA isolated from a human breast cancer specimen, were co-transfected into L40 yeast cells with an S100A1 cDNA bait vector. Target plasmids were recovered from six independently isolated blue colonies, and the purified plasmid DNAs from each colony were co-transfected into strain L40 yeast cells. Reisolation of plasmid DNA and co-transfection were repeated once more. wangg@liv.ac.uk A cDNA library constructed from mRNA isolated from a human breast cancer specimen was used. Biochem. J. (0264-6021) 2004 wang-2004d-1 Heterodimeric interaction and interfaces of S100A1 and S100P. yeasx Saccharomyces cerevisiae (Baker's yeast) 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Wang G., Zhang S., Fernig DG., Spiller D., Martin-Fernandez M., Zhang H., Ding Y., Rao Z., Rudland PS., Barraclough R. Cancer - Interactions investigated in the context of cancer More than 2 E6 yeast cell transformants from a yeast two hybrid cDNA library, constructed from mRNA isolated from a human breast cancer specimen, were co-transfected into L40 yeast cells with an S100A1 cDNA bait vector. Target plasmids were recovered from six independently isolated blue colonies, and the purified plasmid DNAs from each colony were co-transfected into strain L40 yeast cells. Reisolation of plasmid DNA and co-transfection were repeated once more. wangg@liv.ac.uk A cDNA library constructed from mRNA isolated from a human breast cancer specimen was used. Biochem. J. (0264-6021) 2004 wang-2004d-3 Heterodimeric interaction and interfaces of S100A1 and S100P. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Wang G., Zhang S., Fernig DG., Spiller D., Martin-Fernandez M., Zhang H., Ding Y., Rao Z., Rudland PS., Barraclough R. Cancer - Interactions investigated in the context of cancer More than 2 E6 yeast cell transformants from a yeast two hybrid cDNA library, constructed from mRNA isolated from a human breast cancer specimen, were co-transfected into L40 yeast cells with an S100A1 cDNA bait vector. Target plasmids were recovered from six independently isolated blue colonies, and the purified plasmid DNAs from each colony were co-transfected into strain L40 yeast cells. Reisolation of plasmid DNA and co-transfection were repeated once more. wangg@liv.ac.uk A cDNA library constructed from mRNA isolated from a human breast cancer specimen was used. Biochem. J. (0264-6021) 2004 wang-2004d-5 Heterodimeric interaction and interfaces of S100A1 and S100P. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret predetermined predetermined participant predetermined Wang G., Zhang S., Fernig DG., Spiller D., Martin-Fernandez M., Zhang H., Ding Y., Rao Z., Rudland PS., Barraclough R. Cancer - Interactions investigated in the context of cancer More than 2 E6 yeast cell transformants from a yeast two hybrid cDNA library, constructed from mRNA isolated from a human breast cancer specimen, were co-transfected into L40 yeast cells with an S100A1 cDNA bait vector. Target plasmids were recovered from six independently isolated blue colonies, and the purified plasmid DNAs from each colony were co-transfected into strain L40 yeast cells. Reisolation of plasmid DNA and co-transfection were repeated once more. wangg@liv.ac.uk A cDNA library constructed from mRNA isolated from a human breast cancer specimen was used. Biochem. J. (0264-6021) 2004 wang-2004d-4 Heterodimeric interaction and interfaces of S100A1 and S100P. human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells confocal microscopy confocal microscopy predetermined predetermined participant predetermined Wang G., Zhang S., Fernig DG., Spiller D., Martin-Fernandez M., Zhang H., Ding Y., Rao Z., Rudland PS., Barraclough R. Cancer - Interactions investigated in the context of cancer More than 2 E6 yeast cell transformants from a yeast two hybrid cDNA library, constructed from mRNA isolated from a human breast cancer specimen, were co-transfected into L40 yeast cells with an S100A1 cDNA bait vector. Target plasmids were recovered from six independently isolated blue colonies, and the purified plasmid DNAs from each colony were co-transfected into strain L40 yeast cells. Reisolation of plasmid DNA and co-transfection were repeated once more. wangg@liv.ac.uk A cDNA library constructed from mRNA isolated from a human breast cancer specimen was used. Biochem. J. (0264-6021) 2004 s100p_human Protein S100-P S100 calcium-binding protein P S100P S100E Protein S100-E Migration-inducing gene 9 protein protein protein human Homo sapiens Human MTELETAMGMIIDVFSRYSGSEGSTQTLTKGELKVLMEKELPGFLQSGKDKDAVDKLLKDLDANGDAQVDFSEFIVFVAAITSACHKYFEKAGLK s10a1_human Protein S100-A1 S100 calcium-binding protein A1 S-100 protein subunit alpha S-100 protein alpha chain S100A1 S100A protein protein human Homo sapiens Human MGSELETAMETLINVFHAHSGKEGDKYKLSKKELKELLQTELSGFLDAQKDVDAVDKVMKELDENGDGEVDFQEYVVLVAALTVACNNFFWENS myh9_human Myosin-9 MYH9 Myosin heavy chain 9 Myosin heavy chain, non-muscle IIa Non-muscle myosin heavy chain IIa Cellular myosin heavy chain, type A Non-muscle myosin heavy chain A protein protein human Homo sapiens Human MAQQAADKYLYVDKNFINNPLAQADWAAKKLVWVPSDKSGFEPASLKEEVGEEAIVELVENGKKVKVNKDDIQKMNPPKFSKVEDMAELTCLNEASVLHNLKERYYSGLIYTYSGLFCVVINPYKNLPIYSEEIVEMYKGKKRHEMPPHIYAITDTAYRSMMQDREDQSILCTGESGAGKTENTKKVIQYLAYVASSHKSKKDQGELERQLLQANPILEAFGNAKTVKNDNSSRFGKFIRINFDVNGYIVGANIETYLLEKSRAIRQAKEERTFHIFYYLLSGAGEHLKTDLLLEPYNKYRFLSNGHVTIPGQQDKDMFQETMEAMRIMGIPEEEQMGLLRVISGVLQLGNIVFKKERNTDQASMPDNTAAQKVSHLLGINVTDFTRGILTPRIKVGRDYVQKAQTKEQADFAIEALAKATYERMFRWLVLRINKALDKTKRQGASFIGILDIAGFEIFDLNSFEQLCINYTNEKLQQLFNHTMFILEQEEYQREGIEWNFIDFGLDLQPCIDLIEKPAGPPGILALLDEECWFPKATDKSFVEKVMQEQGTHPKFQKPKQLKDKADFCIIHYAGKVDYKADEWLMKNMDPLNDNIATLLHQSSDKFVSELWKDVDRIIGLDQVAGMSETALPGAFKTRKGMFRTVGQLYKEQLAKLMATLRNTNPNFVRCIIPNHEKKAGKLDPHLVLDQLRCNGVLEGIRICRQGFPNRVVFQEFRQRYEILTPNSIPKGFMDGKQACVLMIKALELDSNLYRIGQSKVFFRAGVLAHLEEERDLKITDVIIGFQACCRGYLARKAFAKRQQQLTAMKVLQRNCAAYLKLRNWQWWRLFTKVKPLLQVSRQEEEMMAKEEELVKVREKQLAAENRLTEMETLQSQLMAEKLQLQEQLQAETELCAEAEELRARLTAKKQELEEICHDLEARVEEEEERCQHLQAEKKKMQQNIQELEEQLEEEESARQKLQLEKVTTEAKLKKLEEEQIILEDQNCKLAKEKKLLEDRIAEFTTNLTEEEEKSKSLAKLKNKHEAMITDLEERLRREEKQRQELEKTRRKLEGDSTDLSDQIAELQAQIAELKMQLAKKEEELQAALARVEEEAAQKNMALKKIRELESQISELQEDLESERASRNKAEKQKRDLGEELEALKTELEDTLDSTAAQQELRSKREQEVNILKKTLEEEAKTHEAQIQEMRQKHSQAVEELAEQLEQTKRVKANLEKAKQTLENERGELANEVKVLLQGKGDSEHKRKKVEAQLQELQVKFNEGERVRTELADKVTKLQVELDNVTGLLSQSDSKSSKLTKDFSALESQLQDTQELLQEENRQKLSLSTKLKQVEDEKNSFREQLEEEEEAKHNLEKQIATLHAQVADMKKKMEDSVGCLETAEEVKRKLQKDLEGLSQRHEEKVAAYDKLEKTKTRLQQELDDLLVDLDHQRQSACNLEKKQKKFDQLLAEEKTISAKYAEERDRAEAEAREKETKALSLARALEEAMEQKAELERLNKQFRTEMEDLMSSKDDVGKSVHELEKSKRALEQQVEEMKTQLEELEDELQATEDAKLRLEVNLQAMKAQFERDLQGRDEQSEEKKKQLVRQVREMEAELEDERKQRSMAVAARKKLEMDLKDLEAHIDSANKNRDEAIKQLRKLQAQMKDCMRELDDTRASREEILAQAKENEKKLKSMEAEMIQLQEELAAAERAKRQAQQERDELADEIANSSGKGALALEEKRRLEARIAQLEEELEEEQGNTELINDRLKKANLQIDQINTDLNLERSHAQKNENARQQLERQNKELKVKLQEMEGTVKSKYKASITALEAKIAQLEEQLDNETKERQAACKQVRRTEKKLKDVLLQVDDERRNAEQYKDQADKASTRLKQLKRQLEEAEEEAQRANASRRKLQRELEDATETADAMNREVSSLKNKLRRGDLPFVVPRRMARKGAGDGSDEEVDGKADGAEAKPAE s100a1-s100p-2 545949 N/A 545954 unspecified role unspecified role bait bait N/A 545955 unspecified role unspecified role prey prey P23297:p.[Leu12His;Phe16Ala;Phe72Leu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P23297:p.Phe16Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 1 B, 6 s100p-s100p 545949 N/A 545954 unspecified role unspecified role prey prey P25815:p.[Ile11Ala;Ile12Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe15Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe71Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe74Gly mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe74Val mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Ile75Gly mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Val76Gly mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.[Ile75Gly;Val76Gly] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 545954 unspecified role unspecified role bait bait physical association physical association false false false 1 B, 6 s100a1-s100p-7 545949 N/A 545954 unspecified role unspecified role prey prey P25815:p.[Ile11Ala;Ile12Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe15Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe71Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe74Gly mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Phe74Val mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.Ile75Gly mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P25815:p.[Ile75Gly;Val76Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 545955 unspecified role unspecified role bait bait physical association physical association false false false 1 B, 6 These results suggest that the interface for the S100P-S100A1 interaction is similar but not identical to the S100P-S100P interface. s100a1-s100p-1 545950 N/A 545954 unspecified role unspecified role prey prey N/A 545955 unspecified role unspecified role bait bait physical association physical association false false false 1 A s100a1-s100a1-4 545950 N/A 545955 unspecified role unspecified role bait bait N/A 545955 unspecified role unspecified role prey prey physical association physical association false false false 1 A s100a1-s100p-3 545951 N/A 545954 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) N/A 545955 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false The presence of Ca2+ increases the association rate constant. In the absence of Ca2+, the association rate constant was 30-50-fold slower. Also the Kb is lower in the presence of Ca2+. The Kd in the presence of Ca2+ is 1.4E-8 but in the absence of Ca2+ it is 1.1E-6. 2 E-H, Table 1 myh9-s100p 545951 N/A 545954 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) N/A 545956 unspecified role unspecified role bait bait ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false rhS100A1 has an inhibitory effect on the binding of rhS100P to the immobilized rhC-MHC-IIA. 5 s100a1-s100p-5 545952 N/A 545955 unspecified role unspecified role fluorescence donor fluorescence donor n-terminal ecfp tag ECFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 545954 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept c-terminal eyfp tag EYFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Table 2 s100a1-s100a1-5 545952 N/A 545955 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept c-terminal EYFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 545955 unspecified role unspecified role fluorescence donor fluorescence donor n-terminal ECFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Table 2 s100a1-s100p-4 545953 N/A 545955 unspecified role unspecified role neutral component neutral component n-terminal ECFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 545954 unspecified role unspecified role neutral component neutral component c-terminal EYFP tag fluorescent prot tag fluorescent protein tag fluorescent prot tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false 4 A, B, D The images show whole cells and the colocalization appears to be throughout the whole cell and not localized to any specific region.