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/ FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions rat-heart Rattus norvegicus heart heart Heart [Cardiac; Cardiac tissue] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu Accepted 2008-MAY-1 by DAVET. 14-May-2008: Contacted by JYOTI. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions imex curation FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions 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 Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu Accepted 2008-MAY-1 by DAVET. 14-May-2008: Contacted by JYOTI. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions imex curation FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu Accepted 2008-MAY-16 by DAVET. 14-May-2008: Contacted by JYOTI. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions imex curation FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions 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 Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu Accepted 2008-MAY-1 by DAVET. 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions imex curation FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells 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 Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu Accepted 2008-MAY-1 by DAVET. 14-May-2008: Contacted by JYOTI. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions imex curation FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions rat-heart Rattus norvegicus heart heart Heart [Cardiac; Cardiac tissue] fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Sun J., Yan G., Ren A., You B., Liao JK. add go term for cytoplasm to interaction when go working sunj1@umdnj.edu Accepted 2008-MAY-1 by DAVET. 14-May-2008: Contacted by JYOTI. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions imex curation FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions mouse-heart Mus musculus heart [Cardiac; Cardiac tissue] heart Heart [Cardiac; Cardiac tissue] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sun J., Yan G., Ren A., You B., Liao JK. check again when SwissProt entry for mouse sphingosine kinase 1 is available sunj1@umdnj.edu Accepted 2008-MAY-1 by DAVET. 14-May-2008: Contacted by JYOTI. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions imex curation FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. FHL2/SLIM3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1. Circ. Res. (0009-7330) 2006 imex curation Sun J., Yan G., Ren A., You B., Liao JK. sunj1@umdnj.edu 14-May-2008: Contacted by JYOTI. A human heart cDNA library was used. davet Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Sun J., Yan G., Ren A., You B., Liao JK. add go term for cytoplasm to interaction when go working sunj1@umdnj.edu Accepted 2008-MAY-1 by DAVET. 14-May-2008: Contacted by JYOTI. Circ. Res. (0009-7330) 2006 davet Only protein-protein interactions 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 sphk1_rat Sphingosine kinase 1 Sphk1 Acetyltransferase SPHK1 protein protein rat Rattus norvegicus (Rat) Rat MQPADCPRGLLPRPCRVLVLLNPRGGKGKALKLFQSRVRPLLEEAEVSFKLMLTERQNHARELVCAEELGHWDALAVMSGDGLMHEVVNGLMERPDWESAIQKPLCSLPGGSGNALAASLNYYAGHEQVTNEDLLINCTLLLCCRQLSPMNLLSLHTASGRQLYSVLSLSWGFVADVDLESEKYRSLGEIRFTVGTFFRLASLRIYQGQLAYLPVGKAASKIPASSLAQKGPANTYLVPLEEPVPPHWTVVPEQDFVLVLVLLHTHLSTEMFAAPMGRCEAGVMHLFYIRAGVSRAMLVRLFLAMQKGKHMDLDCPYLVHVPVVAFRLEPRNQRGVFSVDGELMVCEAVQGQVHPNYLWMVSGSSDSPSGRDSQRRPPPEEPI CAAD5817BF4B9507 fhl2_rat Four and a half LIM domains protein 2 Fhl2 Dral Slim3 Skeletal muscle LIM-protein 3 LIM domain protein DRAL protein protein rat Rattus norvegicus (Rat) Rat MTERFDCHHCNESLYGKKYILKEENPHCVACFEELYANTCEECGTPIGCDCKDLSYKDRHWHEGCFHCSRCGSSLVDKPFAAKEEQLLCTDCYSNEYSSKCQECKKTIMPGTRKMEYKGSSWHETCFTCQRCQQPIGTKSFIPKENQNFCVPCYEKQYALQCVQCKKPITTGGVTYRDQPWHRECFVCTACKKQLSGQRFTARDEFPYCLTCFCDLYAKKCAGCTNPISGLGGTKYISFEERQWHNDCFNCKKCSLSLVGRGFLTERDDILCPDCGKDI 9A9D8E5935034173 sphk1_mouse Sphingosine kinase 1 Acetyltransferase SPHK1 Sk1 Sphk1 protein protein mouse Mus musculus Mouse MEPVECPRGLLPRPCRVLVLLNPQGGKGKALQLFQSRVQPFLEEAEITFKLILTERKNHARELVCAEELGHWDALAVMSGDGLMHEVVNGLMERPDWETAIQKPLCSLPGGSGNALAASVNHYAGYEQVTNEDLLINCTLLLCRRRLSPMNLLSLHTASGLRLYSVLSLSWGFVADVDLESEKYRRLGEIRFTVGTFFRLASLRIYQGQLAYLPVGTVASKRPASTLVQKGPVDTHLVPLEEPVPSHWTVVPEQDFVLVLVLLHTHLSSELFAAPMGRCEAGVMHLFYVRAGVSRAALLRLFLAMQKGKHMELDCPYLVHVPVVAFRLEPRSQRGVFSVDGELMVCEAVQGQVHPNYLWMVCGSRDAPSGRDSRRGPPPEEP B791FAA58FCE3D29 sphk2_human Sphingosine kinase 2 SPHK2 SK2 protein protein human Homo sapiens Human MNGHLEAEEQQDQRPDQELTGSWGHGPRSTLVRAKAMAPPPPPLAASTPLLHGEFGSYPARGPRFALTLTSQALHIQRLRPKPEARPRGGLVPLAEVSGCCTLRSRSPSDSAAYFCIYTYPRGRRGARRRATRTFRADGAATYEENRAEAQRWATALTCLLRGLPLPGDGEITPDLLPRPPRLLLLVNPFGGRGLAWQWCKNHVLPMISEAGLSFNLIQTERQNHARELVQGLSLSEWDGIVTVSGDGLLHEVLNGLLDRPDWEEAVKMPVGILPCGSGNALAGAVNQHGGFEPALGLDLLLNCSLLLCRGGGHPLDLLSVTLASGSRCFSFLSVAWGFVSDVDIQSERFRALGSARFTLGTVLGLATLHTYRGRLSYLPATVEPASPTPAHSLPRAKSELTLTPDPAPPMAHSPLHRSVSDLPLPLPQPALASPGSPEPLPILSLNGGGPELAGDWGGAGDAPLSPDPLLSSPPGSPKAALHSPVSEGAPVIPPSSGLPLPTPDARVGASTCGPPDHLLPPLGTPLPPDWVTLEGDFVLMLAISPSHLGADLVAAPHARFDDGLVHLCWVRSGISRAALLRLFLAMERGSHFSLGCPQLGYAAARAFRLEPLTPRGVLTVDGEQVEYGPLQAQMHPGIGTLLTGPPGCPGREP F73FFCEC930DA50F sphk1_human Sphingosine kinase 1 Acetyltransferase SPHK1 SK1 SPHK SPHK1 SPK protein protein human Homo sapiens Human MDPAGGPRGVLPRPCRVLVLLNPRGGKGKALQLFRSHVQPLLAEAEISFTLMLTERRNHARELVRSEELGRWDALVVMSGDGLMHEVVNGLMERPDWETAIQKPLCSLPAGSGNALAASLNHYAGYEQVTNEDLLTNCTLLLCRRLLSPMNLLSLHTASGLRLFSVLSLAWGFIADVDLESEKYRRLGEMRFTLGTFLRLAALRTYRGRLAYLPVGRVGSKTPASPVVVQQGPVDAHLVPLEEPVPSHWTVVPDEDFVLVLALLHSHLGSEMFAAPMGRCAAGVMHLFYVRAGVSRAMLLRLFLAMEKGRHMEYECPYLVYVPVVAFRLEPKDGKGVFAVDGELMVSEAVQGQVHPNYFWMVSGCVEPPPSWKPQQMPPPEEPL EB04A7F2034C2DB0 fhl2_mouse Four and a half LIM domains protein 2 Fhl2 Skeletal muscle LIM-protein 3 protein protein mouse Mus musculus Mouse MTERFDCHHCNESLYGKKYILKEENPHCVACFEELYANTCEECGTPIGCDCKDLSYKDRHWHEGCFHCSRCGSSLVDKPFAAKEEQLLCTDCYSNEYSSKCQECKKTIMPGTRKMEYKGSSWHETCFTCQRCQQPIGTKSFIPKENQNFCVPCYEKQYALQCVQCKKPITTGGVTYREQPWHKECFVCTACKKQLSGQRFTARDEFPYCLTCFCDLYAKKCAGCTNPISGLGGTKYISFEERQWHNDCFNCKKCSLSLVGRGFLTERDDILCPDCGKDI 6D8CBC4B4424BFF2 sphk1-fhl2 4 14 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association a yeast 2-hybrid screening was performed with human SK1 as bait in conjunction with a human heart cDNA library. After screening 2.4 106 clones, a total of 15 positive clones were identified, 3 of which encoded a full-length cDNA of the cardiacrestricted LIM-only factor, FHL2. sphk1-fhl2-1 2 14 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association Coexpression of pGAD-FHL2 and pGBD-SK1 resulted in a 5.1-fold increase in -galactosidase activity compared with coexpression of pGBD and pGAD-FHL2 1 A, C, D sphk1-fhl2-2 5 14 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 1 B cotransfection of the vectors encoding Gal4 DBD-SK1 with FHL2-VP16 resulted in a substantial increase in Gal4-dependent transcriptional activity, indicating interaction of SK1 and FHL2 in mammalian cells. sphk1-fhl2-3 3 14 unspecified role unspecified role bait bait region 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 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 2 A, C Immunoprecipitation of FLAG-tagged SK1 led to coimmunoprecipitation of Myc-tagged FHL2 when both proteins were cotransfected Q14192; Q9NYA1 transfection fhl2-sphk1 3 14 unspecified role unspecified role prey prey region 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 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 2 A, 4 Q14192; Q9NYA1 transfection immunoprecipitation of Myc-tagged FHL2 resulted in coimmunoprecipitation of FLAG-tagged SK1 sphk2-fhl2 3 13 unspecified role unspecified role bait bait region 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 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 2 C Q14192; Q6X2Y2 transfection SK2, which shares approximately 80% similarity with SK1 in the C-terminal portion,11 also interacted with FHL2 sphk1-fhl2-4 7 15 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey physical association physical association 2 D we performed immunoprecipitation with anti-FHL2 antibody using lysates obtained from murine heart. SK1 coprecipitated with the anti-FHL2 antibody fhl2-sphk1-1 8 9 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 14 unspecified role unspecified role neutral component neutral component region 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 3 A Q14192; Q9NYA1 transfection Immunofluorescent microscopy showed that FHL2 and SK1 are colocalized in the cytoplasm sphk11-fhl2-5 6 11 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component colocalization colocalization 3 B Immunofluorescent microscopy showed that FHL2 and SK1 are colocalized in the cytoplasm sphk11-fhl2-6 1 11 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 6 A, 7 A We found that treatment of neonatal rat cardiomyocytes with ET-1 (100 nmol/L, 30 minutes) decreased FHL2 interaction with SK1 Endothelin-1 (ET-1) - Treatment with ET-1 (100 nmol/L, 30 minutes) decreased FHL2 interaction with SK1. siRNA against FHL2 reduced the amount of FHL2 coimmunoprecipitated with SK1. fhl2-sphk1-2 3 14 unspecified role unspecified role prey prey region 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 c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 4 Q14192; Q9NYA1 transfection FHL2 interacted only with SK1 C-terminal portion consisting of C4 and C5 domains