bhf-ucl maruoka-2000-1 alpha-actinin-2 couples to cardiac Kv1.5 channels, regulating current density and channel localization in HEK cells. 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 Maruoka N.D., Steele D.F., Au B.P., Dan P., Zhang X., Moore E.D., Fedida D. Cardiac - Interactions involving cardiac related proteins Source of species information: In Abstract: Human Kv1.5 interacts via its N-terminus/core region and can be immunoprecipitated with α-actinin-2 both after in vitro translation and from HEK cells expressing both proteins. In methods 2.1: screened a 'human cardiac cDNA library' which identified α-actinin-2 2.3: full length α-actinin-2 cloned from 'human cardiac polyA+mRNA' fedida@interchange.ubc.ca A large fragment of Kv1.5 (amino acids (AA) 71-561) was subcloned in frame into the GAL4 DNA binding domain (BD) vector pGBD-C2 and used to screen pGAD10 human cardiac cDNA library (Clontech). FEBS Lett. (0014-5793) 2000 imex curation Only protein-protein interactions maruoka-2000-2 alpha-actinin-2 couples to cardiac Kv1.5 channels, regulating current density and channel localization in HEK cells. 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 Maruoka N.D., Steele D.F., Au B.P., Dan P., Zhang X., Moore E.D., Fedida D. Cardiac - Interactions involving cardiac related proteins Source of species information: In Abstract: Human Kv1.5 interacts via its N-terminus/core region and can be immunoprecipitated with α-actinin-2 both after in vitro translation and from HEK cells expressing both proteins. In methods 2.1: screened a 'human cardiac cDNA library' which identified α-actinin-2 2.3: full length α-actinin-2 cloned from 'human cardiac polyA+mRNA' fedida@interchange.ubc.ca A large fragment of Kv1.5 (amino acids (AA) 71-561) was subcloned in frame into the GAL4 DNA binding domain (BD) vector pGBD-C2 and used to screen pGAD10 human cardiac cDNA library (Clontech). FEBS Lett. (0014-5793) 2000 imex curation Only protein-protein interactions maruoka-2000-3 alpha-actinin-2 couples to cardiac Kv1.5 channels, regulating current density and channel localization in HEK cells. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Maruoka N.D., Steele D.F., Au B.P., Dan P., Zhang X., Moore E.D., Fedida D. Cardiac - Interactions involving cardiac related proteins Source of species information: In Abstract: Human Kv1.5 interacts via its N-terminus/core region and can be immunoprecipitated with α-actinin-2 both after in vitro translation and from HEK cells expressing both proteins. In methods 2.1: screened a 'human cardiac cDNA library' which identified α-actinin-2 2.3: full length α-actinin-2 cloned from 'human cardiac polyA+mRNA' fedida@interchange.ubc.ca A large fragment of Kv1.5 (amino acids (AA) 71-561) was subcloned in frame into the GAL4 DNA binding domain (BD) vector pGBD-C2 and used to screen pGAD10 human cardiac cDNA library (Clontech). FEBS Lett. (0014-5793) 2000 imex curation Only protein-protein interactions maruoka-2000-4 alpha-actinin-2 couples to cardiac Kv1.5 channels, regulating current density and channel localization in HEK cells. 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 bait coip anti bait coimmunoprecipitation anti bait coip polyclonal western polyclonal antibody western blot polyclonal western Maruoka N.D., Steele D.F., Au B.P., Dan P., Zhang X., Moore E.D., Fedida D. Cardiac - Interactions involving cardiac related proteins Source of species information: In Abstract: Human Kv1.5 interacts via its N-terminus/core region and can be immunoprecipitated with α-actinin-2 both after in vitro translation and from HEK cells expressing both proteins. In methods 2.1: screened a 'human cardiac cDNA library' which identified α-actinin-2 2.3: full length α-actinin-2 cloned from 'human cardiac polyA+mRNA' fedida@interchange.ubc.ca A large fragment of Kv1.5 (amino acids (AA) 71-561) was subcloned in frame into the GAL4 DNA binding domain (BD) vector pGBD-C2 and used to screen pGAD10 human cardiac cDNA library (Clontech). FEBS Lett. (0014-5793) 2000 imex curation Only protein-protein interactions maruoka-2000-5 alpha-actinin-2 couples to cardiac Kv1.5 channels, regulating current density and channel localization in HEK cells. 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 bait coip anti bait coimmunoprecipitation anti bait coip monoclonal western monoclonal antibody western blot monoclonal western Maruoka N.D., Steele D.F., Au B.P., Dan P., Zhang X., Moore E.D., Fedida D. Cardiac - Interactions involving cardiac related proteins Source of species information: In Abstract: Human Kv1.5 interacts via its N-terminus/core region and can be immunoprecipitated with α-actinin-2 both after in vitro translation and from HEK cells expressing both proteins. In methods 2.1: screened a 'human cardiac cDNA library' which identified α-actinin-2 2.3: full length α-actinin-2 cloned from 'human cardiac polyA+mRNA' fedida@interchange.ubc.ca A large fragment of Kv1.5 (amino acids (AA) 71-561) was subcloned in frame into the GAL4 DNA binding domain (BD) vector pGBD-C2 and used to screen pGAD10 human cardiac cDNA library (Clontech). FEBS Lett. (0014-5793) 2000 imex curation Only protein-protein interactions kcna5_human Potassium voltage-gated channel subfamily A member 5 KCNA5 HPCN1 Voltage-gated potassium channel HK2 Voltage-gated potassium channel subunit Kv1.5 protein protein human Homo sapiens Human MEIALVPLENGGAMTVRGGDEARAGCGQATGGELQCPPTAGLSDGPKEPAPKGRGAQRDADSGVRPLPPLPDPGVRPLPPLPEELPRPRRPPPEDEEEEGDPGLGTVEDQALGTASLHHQRVHINISGLRFETQLGTLAQFPNTLLGDPAKRLRYFDPLRNEYFFDRNRPSFDGILYYYQSGGRLRRPVNVSLDVFADEIRFYQLGDEAMERFREDEGFIKEEEKPLPRNEFQRQVWLIFEYPESSGSARAIAIVSVLVILISIITFCLETLPEFRDERELLRHPPAPHQPPAPAPGANGSGVMAPPSGPTVAPLLPRTLADPFFIVETTCVIWFTFELLVRFFACPSKAGFSRNIMNIIDVVAIFPYFITLGTELAEQQPGGGGGGQNGQQAMSLAILRVIRLVRVFRIFKLSRHSKGLQILGKTLQASMRELGLLIFFLFIGVILFSSAVYFAEADNQGTHFSSIPDAFWWAVVTMTTVGYGDMRPITVGGKIVGSLCAIAGVLTIALPVPVIVSNFNYFYHRETDHEEPAVLKEEQGTQSQGPGLDRGVQRKVSGSRGSFCKAGGTLENADSARRGSCPLEKCNVKAKSNVDLRRSLYALCLDTSRETDL actn2_human Alpha-actinin-2 ACTN2 Alpha-actinin skeletal muscle isoform 2 F-actin cross-linking protein protein protein human Homo sapiens Human MNQIEPGVQYNYVYDEDEYMIQEEEWDRDLLLDPAWEKQQRKTFTAWCNSHLRKAGTQIENIEEDFRNGLKLMLLLEVISGERLPKPDRGKMRFHKIANVNKALDYIASKGVKLVSIGAEEIVDGNVKMTLGMIWTIILRFAIQDISVEETSAKEGLLLWCQRKTAPYRNVNIQNFHTSWKDGLGLCALIHRHRPDLIDYSKLNKDDPIGNINLAMEIAEKHLDIPKMLDAEDIVNTPKPDERAIMTYVSCFYHAFAGAEQAETAANRICKVLAVNQENERLMEEYERLASELLEWIRRTIPWLENRTPEKTMQAMQKKLEDFRDYRRKHKPPKVQEKCQLEINFNTLQTKLRISNRPAFMPSEGKMVSDIAGAWQRLEQAEKGYEEWLLNEIRRLERLEHLAEKFRQKASTHETWAYGKEQILLQKDYESASLTEVRALLRKHEAFESDLAAHQDRVEQIAAIAQELNELDYHDAVNVNDRCQKICDQWDRLGTLTQKRREALERMEKLLETIDQLHLEFAKRAAPFNNWMEGAMEDLQDMFIVHSIEEIQSLITAHEQFKATLPEADGERQSIMAIQNEVEKVIQSYNIRISSSNPYSTVTMDELRTKWDKVKQLVPIRDQSLQEELARQHANERLRRQFAAQANAIGPWIQNKMEEIARSSIQITGALEDQMNQLKQYEHNIINYKNNIDKLEGDHQLIQEALVFDNKHTNYTMEHIRVGWELLLTTIARTINEVETQILTRDAKGITQEQMNEFRASFNHFDRRKNGLMDHEDFRACLISMGYDLGEAEFARIMTLVDPNGQGTVTFQSFIDFMTRETADTDTAEQVIASFRILASDKPYILAEELRRELPPDQAQYCIKRMPAYSGPGSVPGALDYAAFSSALYGESDL actn2-kcna5 1436122 n/a Kv1.5 1436127 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a alpha-actinin-2 1436128 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1436133 1436131 physical association physical association false false false Fig 1A kcna5-actn2-2 1436123 n/a Kv1.5 1436127 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a alpha-actinin-2 1436128 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1436138 1436136 physical association physical association false false false Fig 1C actn2-kcna5-1 1436124 n/a Kv1.5 1436127 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated 35s 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes n/a alpha-actinin-2 1436128 unspecified role unspecified role bait bait in vitro translated in vitro translated protein in vitro translated rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes direct interaction direct interaction false false false Fig 2A actn2-kcna5-2 1436125 n/a Kv1.5 1436127 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated n/a alpha-actinin-2 1436128 unspecified role unspecified role bait bait in vitro translated in vitro translated protein in vitro translated physical association physical association false false false Fig 2B kcna5-actn2-3 1436126 n/a Kv1.5 1436127 unspecified role unspecified role bait bait in vitro translated in vitro translated protein in vitro translated n/a alpha-actinin-2 1436128 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated physical association physical association false false false Fig 2C