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 Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. The Atypical Histone MacroH2A1.2 Interacts with HER-2 Protein in Cancer Cells. J. Biol. Chem. (0021-9258) 2012 imex curation Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Li X., Kuang J., Shen Y., Majer M.M., Nelson C.C., Parsawar K., Heichman K.A., Kuwada S.K. Cancer - Interactions investigated in the context of cancer This experiment relates to a negative interaction. The "MYC epitope-tagged mH2A1.1 and mH2A1.2 constructs" have been confirmed to be of human origin by the authors. Similarly MYC and V5 tagged constucts have been confirmed as having an additional His tag; Figure 4B - antibodies were used for detection of participants, and Figure 8G uses SKOV-3 cells, not 293t cells. Full-length human ERBB2 was subcloned into the tandem affinity purification vector pNTAP (which has two affinity tags:-a streptavidin binding peptide (SBP) and a calmodulin binding peptide (CBP). ), and then transfected into 293T cells. Lysates from these TAP tagged ERBB2 cells were incubated with streptavidin beads and proteins eluted with biotin. The isoform "mH2A1.2" has been assumed to be UniProt O75367-3 (Isoform 3) based on the quoted mutations (E202G/S204A) which are only found in this isoform. skkuwada@hawaii.edu Accepted 2012-AUG-07 by JYOTI 19-Sept-2012: Contacted by IntAct-Help. J. Biol. Chem. (0021-9258) 2012 Only protein-protein interactions imex curation o75367-2 Core histone macro-H2A.1 MACROH2A1 H2AFY Histone H2A.y Medulloblastoma antigen MU-MB-50.205 mH2A1.1 protein protein human Homo sapiens Human MSSRGGKKKSTKTSRSAKAGVIFPVGRMLRYIKKGHPKYRIGVGAPVYMAAVLEYLTAEILELAGNAARDNKKGRVTPRHILLAVANDEELNQLLKGVTIASGGVLPNIHPELLAKKRGSKGKLEAIITPPPAKKAKSPSQKKPVSKKAGGKKGARKSKKKQGEVSKAASADSTTEGTPADGFTVLSTKSLFLGQKLQVVQADIASIDSDAVVHPTNTDFYIGGEVGNTLEKKGGKEFVEAVLELRKKNGPLEVAGAAVSAGHGLPAKFVIHCNSPVWGADKCEELLEKTVKNCLALADDKKLKSIAFPSIGSGRNGFPKQTAAQLILKAISSYFVSTMSSSIKTVYFVLFDSESIGIYVQEMAKLDAN Specifically binds ADP-ribose and O-acetyl-ADP-ribose. Important residues for binding are Asp-203, Gly-224, Gly-314 and Phe-348. Preferentially expressed over isoform 2 in the absence of DDX5/DDX17. D21A4CE6C7AA3BB1 erbb2_human Receptor tyrosine-protein kinase erbB-2 p185erbB2 MLN19 Proto-oncogene c-ErbB-2 Tyrosine kinase-type cell surface receptor HER2 Proto-oncogene Neu ERBB2 HER2 NEU NGL Metastatic lymph node gene 19 protein protein protein human Homo sapiens Human MELAALCRWGLLLALLPPGAASTQVCTGTDMKLRLPASPETHLDMLRHLYQGCQVVQGNLELTYLPTNASLSFLQDIQEVQGYVLIAHNQVRQVPLQRLRIVRGTQLFEDNYALAVLDNGDPLNNTTPVTGASPGGLRELQLRSLTEILKGGVLIQRNPQLCYQDTILWKDIFHKNNQLALTLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPCARVCYGLGMEHLREVRAVTSANIQEFAGCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLGISWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLHTANRPEDECVGEGLACHQLCARGHCWGPGPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLTSIISAVVGILLVVVLGVVFGILIKRRQQKIRKYTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLGPASPLDSTFYRSLLEDDDMGDLVDAEEYLVPQQGFFCPDPAPGAGGMVHHRHRSSSTRSGGGDLTLGLEPSEEEAPRSPLAPSEGAGSDVFDGDLGMGAAKGLQSLPTHDPSPLQRYSEDPTVPLPSETDGYVAPLTCSPQPEYVNQPDVRPQPPSPREGPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLTPQGGAAPQPHPPPAFSPAFDNLYYWDQDPPERGAPPSTFKGTPTAENPEYLGLDVPV DIP protein P04626 original sequence version: 198 DIP protein P04626 original sequence version: 122 DIP protein P04626 original sequence version: 206 DIP protein P04626 original sequence version: 195 DIP protein P04626 original sequence version: 150 DIP protein P04626 original sequence version: 179 DIP protein P04626 original sequence version: 146 DIP protein P04626 original sequence version: 147 39E9DFDA04DCF962 h2afy-erbb2-1 1 2 unspecified role unspecified role bait bait c_terminus 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 c_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 3 unspecified role unspecified role prey prey physical association physical association true Fig 3A