bhf-ucl furukawa-2001-1 Specific interaction of the potassium channel beta-subunit minK with the sarcomeric protein T-cap suggests a T-tubule-myofibril linking system. 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 Furukawa T., Ono Y., Tsuchiya H., Katayama Y., Bang M.L., Labeit D., Labeit S., Inagaki N., Gregorio C.C. Cardiac - Interactions involving cardiac related proteins Subfragments of the coding sequence of minK from human were obtained by PCR amplifications rat heart cDNA library was used to isolate Tcap gregorio@u.arizona.edu rat heart cDNA library in pACT2 vector (Clontech, Palo Alto, CA) J. Mol. Biol. (0022-2836) 2001 imex curation Only protein-protein interactions furukawa-2001-2 Specific interaction of the potassium channel beta-subunit minK with the sarcomeric protein T-cap suggests a T-tubule-myofibril linking system. in vitro In vitro pull down pull down autoradiography autoradiography Furukawa T., Ono Y., Tsuchiya H., Katayama Y., Bang M.L., Labeit D., Labeit S., Inagaki N., Gregorio C.C. Cardiac - Interactions involving cardiac related proteins Subfragments of the coding sequence of minK from human were obtained by PCR amplifications rat heart cDNA library was used to isolate Tcap gregorio@u.arizona.edu rat heart cDNA library in pACT2 vector (Clontech, Palo Alto, CA) J. Mol. Biol. (0022-2836) 2001 imex curation Only protein-protein interactions furukawa-2001-3 Specific interaction of the potassium channel beta-subunit minK with the sarcomeric protein T-cap suggests a T-tubule-myofibril linking system. 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 Furukawa T., Ono Y., Tsuchiya H., Katayama Y., Bang M.L., Labeit D., Labeit S., Inagaki N., Gregorio C.C. Cardiac - Interactions involving cardiac related proteins Subfragments of the coding sequence of minK from human were obtained by PCR amplifications rat heart cDNA library was used to isolate Tcap gregorio@u.arizona.edu rat heart cDNA library in pACT2 vector (Clontech, Palo Alto, CA) J. Mol. Biol. (0022-2836) 2001 imex curation Only protein-protein interactions kcne1_human Potassium voltage-gated channel subfamily E member 1 KCNE1 IKs producing slow voltage-gated potassium channel subunit beta Mink Minimal potassium channel Delayed rectifier potassium channel subunit IsK protein protein human Homo sapiens Human MILSNTTAVTPFLTKLWQETVQQGGNMSGLARRSPRSSDGKLEALYVLMVLGFFGFFTLGIMLSYIRSKKLEHSNDPFNVYIESDAWQEKDKAYVQARVLESYRSCYVVENHLAIEQPNTHLPETKPSP mint a6his0_rat Tcap rCG_32667 protein protein rat Rattus norvegicus (Rat) Rat MATSELSCQVSEENQERREAFWAEWKDLTLSTRPEEGCSLHEEDTQRHETYHRQGQCQAVVQRSPWLVMRLGILGRGLQEYQLPYQRVLPLPIFTPTKVGATKEEREETPIQLRELLALETALGGQCVERQDVAEITKQLPPVVPVSKPGPLRRTLSRSMSQEAQRG kcne1-tcap 1443733 n/a 1443736 unspecified role unspecified role bait bait mink-ct sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1443737 unspecified role unspecified role prey prey c-terminal 16aa sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1443742 1443740 physical association physical association false false false Fig 1b tcap-kcne1 1443734 n/a 1443736 unspecified role unspecified role prey prey cell lysate cell lysate mink-ct sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 35s 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 autoradiography autoradiography undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes n/a 1443737 unspecified role unspecified role bait bait purified purified his6-tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig 3 kcne1-tcap-1 1443735 n/a 1443736 unspecified role unspecified role bait bait mink-ct sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1443737 unspecified role unspecified role prey prey A6HIS0:p.Ser157Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false A6HIS0:p.Ser157Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false A6HIS0:p.Ser157Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false c-terminal 16aa sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal 8aa required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false A6HIS0:p.[Thr155Glu;Ser157Glu;Ser159Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength 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 physical association physical association false false false Fig 2a, 2b, 4b, 4c