MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. Blood cancer journal 2011 imex curation Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de s.bohlander@auckland.ac.nz Accepted 2018-FEB-22 AT 11:56 GMT AT 11:56 GMT by MGT Blood cancer journal 2011 Only protein-protein interactions imex curation imex curation FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. Blood cancer journal 2011 imex curation Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de s.bohlander@auckland.ac.nz Accepted 2018-FEB-22 AT 11:56 GMT AT 11:56 GMT by MGT Blood cancer journal 2011 Only protein-protein interactions imex curation imex curation FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. Blood cancer journal 2011 imex curation Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de s.bohlander@auckland.ac.nz Accepted 2018-FEB-22 AT 11:56 GMT AT 11:56 GMT by MGT Blood cancer journal 2011 Only protein-protein interactions imex curation imex curation FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. Blood cancer journal 2011 imex curation Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference confocal microscopy confocal microscopy immunostaining immunostaining Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de s.bohlander@auckland.ac.nz Accepted 2018-FEB-22 AT 11:56 GMT AT 11:56 GMT by MGT Blood cancer journal 2011 Only protein-protein interactions imex curation imex curation FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. Blood cancer journal 2011 imex curation Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de s.bohlander@auckland.ac.nz Accepted 2018-FEB-22 AT 11:56 GMT AT 11:56 GMT by MGT Blood cancer journal 2011 Only protein-protein interactions imex curation imex curation FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. FHL2 interacts with CALM and is highly expressed in acute erythroid leukemia. Blood cancer journal 2011 imex curation Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Pašaliç Z., Greif PA., Jurinoviç V., Mulaw M., Kakadia PM., Tizazu B., Fröhlich-Archangelo L., Krause A., Bohlander SK. Cancer - Interactions investigated in the context of cancer About the proteins, I wrote to the Author and he answered me that the proteins are all from Homo Sapiens The authors have stated that constructs used were of Human origin. bohlander@helmholtz-muenchen.de s.bohlander@auckland.ac.nz Accepted 2018-FEB-22 AT 11:56 GMT AT 11:56 GMT by MGT Blood cancer journal 2011 Only protein-protein interactions imex curation imex curation fhl2_human Four and a half LIM domains protein 2 Skeletal muscle LIM-protein 3 LIM domain protein DRAL FHL2 DRAL SLIM3 protein protein human Homo sapiens Human MTERFDCHHCNESLFGKKYILREESPYCVVCFETLFANTCEECGKPIGCDCKDLSYKDRHWHEACFHCSQCRNSLVDKPFAAKEDQLLCTDCYSNEYSSKCQECKKTIMPGTRKMEYKGSSWHETCFICHRCQQPIGTKSFIPKDNQNFCVPCYEKQHAMQCVQCKKPITTGGVTYREQPWHKECFVCTACRKQLSGQRFTARDDFAYCLNCFCDLYAKKCAGCTNPISGLGGTKYISFEERQWHNDCFNCKKCSLSLVGRGFLTERDDILCPDCGKDI DIP protein Q14192 original sequence version: null DIP protein Q14192 original sequence version: 142 DIP protein Q14192 original sequence version: 136 E94E3A7F6601F9F3 pical_human Phosphatidylinositol-binding clathrin assembly protein CALM Clathrin assembly lymphoid myeloid leukemia protein PICALM protein protein human Homo sapiens Human MSGQSLTDRITAAQHSVTGSAVSKTVCKATTHEIMGPKKKHLDYLIQCTNEMNVNIPQLADSLFERTTNSSWVVVFKSLITTHHLMVYGNERFIQYLASRNTLFNLSNFLDKSGLQGYDMSTFIRRYSRYLNEKAVSYRQVAFDFTKVKRGADGVMRTMNTEKLLKTVPIIQNQMDALLDFNVNSNELTNGVINAAFMLLFKDAIRLFAAYNEGIINLLEKYFDMKKNQCKEGLDIYKKFLTRMTRISEFLKVAEQVGIDRGDIPDLSQAPSSLLDALEQHLASLEGKKIKDSTAASRATTLSNAVSSLASTGLSLTKVDEREKQAALEEEQARLKALKEQRLKELAKKPHTSLTTAASPVSTSAGGIMTAPAIDIFSTPSSSNSTSKLPNDLLDLQQPTFHPSVHPMSTASQVASTWGDPFSATVDAVDDAIPSLNPFLTKSSGDVHLSISSDVSTFTTRTPTHEMFVGFTPSPVAQPHPSAGLNVDFESVFGNKSTNVIVDSGGFDELGGLLKPTVASQNQNLPVAKLPPSKLVSDDLDSSLANLVGNLGIGNGTTKNDVNWSQPGEKKLTGGSNWQPKVAPTTAWNAATMAPPVMAYPATTPTGMIGYGIPPQMGSVPVMTQPTLIYSQPVMRPPNPFGPVSGAQIQFM AC3227E9D32AFEDA calm/af10_fusion CALM/AF10 (MLLT10) fusion protein protein protein human Homo sapiens Human MSGQSLTDRITAAQHSVTGSAVSKTVCKATTHEIMGPKKKHLDYLIQCTNEMNVNIPQLADSLFERTTNSSWVVVFKSLITTHHLMVYGNERFIQYLASRNTLFNLSNFLDKSGLQGYDMSTFIRRYSRYLNEKAVSYRQVAFDFTKVKRGADGVMRTMNTEKLLKTVPIIQNQMDALLDFNVNSNELTNGVINAAFMLLFKDAIRLFAAYNEGIINLLEKYFDMKKNQCKEGLDIYKKFLTRMTRISEFLKVAEQVGIDRGDIPDLSQAPSSLLDALEQHLASLEGKKIKDSTAASRATTLSNAVSSLASTGLSLTKVDEREKQAALEEEQARLKALKEQRLKELAKKPHTSLTTAASPVSTSAGGIMTAPAIDIFSTPSSSNSTSKLPNDLLDLQQPTFHPSVHPMSTASQVASTWGDPFSATVDAVDDAIPSLNPFLTKSSGDVHLSISSDVSTFTTRTPTHEMFVGFTPSPVAQPHPSAGLNVDFESVFGNKSTNVIVDSGGFDELGGLLKPTVASQNQNLPVAKLPPSKLVSDDLDSSLANLVGNLGIGNGTTKNDVNWSQPGEKKLTGGSNWQPKVAPTTAWNAATMAPPVMAYPATTPTGMIGYGIPPQMGSVPVMTQPTLIYSQPVMRPPNPFGPVSGAQRCELCPHKDGALKRTDNGGWAHVVCALYIPEVQFANVSTMEPIVLQSVPHDRYNKTCYICDEQGRESKAATGACMTCNKHGCRQAFHVTCAQFAGLLCEEEGNGADNVQYCGYCKYHFSKLKKSKRGSNRSYDQSLSDSSSHSQDKHHEKEKKKYKEKDKHKQKHKKQPEPSPALVPSLTVTTEKTYTSTSNNSISGSLKRLEDTTARFTNANFQEVSAHTSSGKDVSETRGSEGKGKKSSAHSSGQRGRKPGGGRNPGTTVSAASPFPQGSFSGTPGSVKSSSGSSVQSPQDFLSFTDSDLRNDSYSHSQQSSATKDVHKGESGSQEGGVNSFSTLIGLPSTSAVTSQPKSFENSPGDLGNSSLPTAGYKRAQTSGIEEETVKEKKRKGNKQSKHGPGRPKGNKNQENVSHLSVSSASPTSSVASAAGSITSSSLQKSPTLLRNGSLQSLSVGSSPVGSEISMQYRHDGACPTTTFSELLNAIHNGIYNSNDVAVSFPNVVSGSGSSTPVSSSHLPQQSSGHLQQVGALSPSAVSSAAPAVATTQANTLSGSSLSQAPSHMYGNRSNSSMAALIAQSENNQTDQDLGDNSRNLVGRGSSPRGSLSPRSPVSSLQIRYDQPGNSSLENLPPVAASIEQLLERQWSEGQQFLLEQGTPSDILGMLKSLHQLQVENRRLEEQIKNLTAKKERLQLLNAQLSVPFPTITANPSPSHQIHTFSAQTAPTTDSLNSSKSPHIGNSFLPDNSLPVLNQDLTSSGQSTSSSSALSTPPPAGQSPAQQGSGVSGVQQVNGVTVGALASGMQPVTSTIPAVSAVGGIIGALPGNQLAINGIVGALNGVMQTPVTMSQNPTPLTHTTVPPNATHPMPATLTNSASGLGLLSDQQRQILIHQQQFQQLLNSQQLTPEQHQAFLYQLMQHHHQQHHQPELQQLQIPGPTQIPINNLLAGTQAPPLHTATTNPFLTIHGDNASQKVARLSDKTGPVAQEKS fhl2-picalm-1 1 8 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated binding region binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 35s radiolabel 35s radiolabel 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 certain certain sequence position certain certain certain sequence position certain in vitro In vitro 7 unspecified role unspecified role bait bait purified purified N-terminal gst tag gst tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction f2a To confirm the interaction between CALM and FHL2 in vitro, a glutathione S-transferase (GST)-pulldown assay was performed. FHL2 was expressed as a GST fusion protein in bacteria and immobilized on glutathione beads. The beads were then incubated with 35S-methionine-labeled CALM protein (aa 1–408). FHL2, but not GST alone, retained the radiolabeled CALM protein. These results indicate a direct physical interaction between CALM and FHL2. fhl2-picalm-2 2 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f2b After transient transfection of FLAG-FHL2- and YFP-CALM-expressing plasmids in HEK293T cells, and immunoprecipitation of YFP-CALM using GFP-binder beads,29 the co-precipitated FHL2 protein could be detected with anti-FLAG antibodies fhl2-picalm-3 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell v5 tag v5 tag v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fs1 We were also able to co-precipitate small amounts of overexpressed FHL2 by immunoprecipitation of endogenous CALM in HEK293T cells fhl2-picalm-4 5 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding region binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association f3b To map the minimal FHL2 interaction domain of CALM, yeast cells were co-transformed with several CALM deletion mutants fused to the GAL4-DNA-binding domain (DBD) and the FHL2 prey clone. Only transformants with CALM mutants containing amino acids 294–335 grew on selective plates indicating protein–protein interaction fhl2-picalm-5 5 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding region binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell physical association physical association f3c To map the minimal CALM interaction domain of FHL2, yeast cells were co-transformed with several FHL2 deletion mutants fused to GAL4-DBD and with a CALM prey clone, obtained in the Y2H screening using the CALM bait. Only transformants with full-length FHL2 grew on selective plates indicating protein–protein interaction. These results show that amino acids 206–279 of FHL2 are required for its interaction with CALM. fhl2-picalm-6 4 7 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tag flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 tag fluorescence tag visualisation by fluorescence tag fluorescence immunostaining immunostaining unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell region yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization f4m, 5A. There was also colocalization of YFP-CALM and FLAG-FHL2 in the nucleus and especially in the cytoplasm (Figures 4j–m) This colocalization can be visualized in line scan images in Figure 5a. calm/af10 fusion-fhl2-1 4 7 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell region flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 tag fluorescence tag visualisation by fluorescence tag fluorescence immunostaining immunostaining unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell region yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization f4q, 5B When FLAG-FHL2 was co-expressed with YFPCALM/ AF10 (Figures 4n–q), the subcellular localization of YFPCALM/ AF10 did not change markedly (compare Figures 4e and o), but there were pronounced changes in the cytoplasmic localization of FLAG-FHL2 (compare Figures 4h and p), which colocalized with YFP-CALM/AF10 in distinct cytoplasmic speckles. The colocalization of FLAG-FHL2 and YFP-CALM/AF10 was also clearly visible in the line scans of the confocal images (Figure 5b). calm/af10 fusion-fhl2-2 4 7 tag fluorescence tag visualisation by fluorescence tag fluorescence immunostaining immunostaining unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n_terminus yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell region flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization f4u, 5C The same pattern of colocalization of CALM/AF10 and FHL2 was also observed when a FLAGtagged CALM/AF10 and YFP-FHL2 fusion protein were coexpressed The colocalization can be visualized in line scan images in figure 5c fhl2-picalm-7 3 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell v5 tag v5 tag v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell yfp tag yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association fs1 Precipitation of YFP-CALM with CALM antibodies co-precipitates V5-FHL2. FHL2 is detected with a V5 antibody. fhl2-picalm-8 6 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain physical association physical association no figure To identify interaction partners of the CALM protein, the N-terminal 408 amino acids of CALM were used as a bait to perform a Y2H screen of a human thymus cDNA library. Under high stringency screening conditions of 1106 primary transformants, six clones were identified that grew in the absence of histidine and adenine and were b-galactosidase positive. These six clones encoded five different proteins. One of these clones contained the complete open reading frame of FHL2