bhf-ucl singh-2013-1 Abnormal calcium cycling and cardiac arrhythmias associated with the human Ser96Ala genetic variant of histidine-rich calcium-binding protein. 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 Singh V.P., Rubinstein J., Arvanitis D.A., Ren X., Gao X., Haghighi K., Gilbert M., Iyer V.R., Kim d.o. .H., Cho C., Jones K., Lorenz J.N., Armstrong C.F., Wang H.S., Gyorke S., Kranias E.G. Cardiac - Interactions involving cardiac related proteins from Methods: Full‐length green fluorescent protein (GFP)‐rabbit triadin (GenBank: U31540) and GFP‐hHRC Ser96 or GFP‐hHRC Ala96 constructs were transiently transfected in HEK 293 cells. human HRC cDNA with the HRC Ser96 or HRC Ala96 mutation. Transgenic C57/BL6 mice with cardiac‐specific expression of the human HRC cDNA carrying either the human Ser96 HRC or Ala96 HRC were mated with the HRC‐KO mice (C57/BL6) F1 heterozygous HRC offspring with the HRC mutant transgenes were identified by using PCR methodology and bred with HRC‐KO mice to obtain the F2 generation. The HRC‐KO offspring carrying the human HRC variant transgenes were selected to backcross with HRC‐KO mice for at least 3 generations before using them for our studies. Litsa.Kranias@uc.edu J Am Heart Assoc 2013 Only protein-protein interactions imex curation singh-2013-2 Abnormal calcium cycling and cardiac arrhythmias associated with the human Ser96Ala genetic variant of histidine-rich calcium-binding protein. 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 anti tag western anti tag western blot anti tag western Singh V.P., Rubinstein J., Arvanitis D.A., Ren X., Gao X., Haghighi K., Gilbert M., Iyer V.R., Kim d.o. .H., Cho C., Jones K., Lorenz J.N., Armstrong C.F., Wang H.S., Gyorke S., Kranias E.G. Cardiac - Interactions involving cardiac related proteins from Methods: Full‐length green fluorescent protein (GFP)‐rabbit triadin (GenBank: U31540) and GFP‐hHRC Ser96 or GFP‐hHRC Ala96 constructs were transiently transfected in HEK 293 cells. human HRC cDNA with the HRC Ser96 or HRC Ala96 mutation. Transgenic C57/BL6 mice with cardiac‐specific expression of the human HRC cDNA carrying either the human Ser96 HRC or Ala96 HRC were mated with the HRC‐KO mice (C57/BL6) F1 heterozygous HRC offspring with the HRC mutant transgenes were identified by using PCR methodology and bred with HRC‐KO mice to obtain the F2 generation. The HRC‐KO offspring carrying the human HRC variant transgenes were selected to backcross with HRC‐KO mice for at least 3 generations before using them for our studies. Litsa.Kranias@uc.edu J Am Heart Assoc 2013 Only protein-protein interactions imex curation trdn_mouse Triadin Trdn protein protein mouse Mus musculus Mouse MTEITAEGNASTTTTVIDNKNGCIPKSPGKVLKRSVTEDIVTTFSSPAAWLLVIALIITWSAVAIVMFDLVDYKNFSASSIAKIGSDPLKLVNDAVEETTDWIYGFFSLLSDIISSEGDEDDEDADEDIDKGEIEEPPLKRKEIHQEKAEKEEKPEKKIQTKASHREREKGKEKLKGEKPEKTATHKEKLEKKERPETKMMAKEDKKIKTKEKTEEKAKKEMKVGKQEKVKPTAAKAKETPKTPPKARKKDDKEMPAVHEQKDQYAFCRYMIDMFVHGDLKPGQSPVMPPPSLTPSKPALSTTALEEKEKEEKKKMEKKDTSDTKKKEKEVKKKSEETTIDGKGKEPGKPPETKQMTAKLTTQAAARKDEKKEESKKMRKPTEEQPKGKKQEKKEKHIEPAKTPKKEHPGPSEKLKKAKAEQAKEEIAAASTKKALHGKKEEKAKTVEQEVKKEKSGKSSSDLKDKEVKKEKSGKSSSDLKDKEPQLKNEEKSKPQVKKEAKLASSDKGQTRKQNITRPEQVIPHVKPEKAEHQEKGHPSIKKDKPKPSSKGAPEVPDSGKKKIEKSEKESKVPTREENLQVYNVTKAEKPGKIPKDSKEAPASKKDKEDSKEAPTSKKDKEDSKDVPHSKKDKEVTDDVSSPKKQTRPISFFQCVYLNGYNGYGFQFPVTPVQQPGENPGKTNSPGQKQQEQ srch_human Sarcoplasmic reticulum histidine-rich calcium-binding protein HRC HCP protein protein human Homo sapiens Human MGHHRPWLHASVLWAGVASLLLPPAMTQQLRGDGLGFRNRNNSTGVAGLSEEASAELRHHLHSPRDHPDENKDVSTENGHHFWSHPDREKEDEDVSKEYGHLLPGHRSQDHKVGDEGVSGEEVFAEHGGQARGHRGHGSEDTEDSAEHRHHLPSHRSHSHQDEDEDEVVSSEHHHHILRHGHRGHDGEDDEGEEEEEEEEEEEEASTEYGHQAHRHRGHGSEEDEDVSDGHHHHGPSHRHQGHEEDDDDDDDDDDDDDDDDVSIEYRHQAHRHQGHGIEEDEDVSDGHHHRDPSHRHRSHEEDDNDDDDVSTEYGHQAHRHQDHRKEEVEAVSGEHHHHVPDHRHQGHRDEEEDEDVSTERWHQGPQHVHHGLVDEEEEEEEITVQFGHYVASHQPRGHKSDEEDFQDEYKTEVPHHHHHRVPREEDEEVSAELGHQAPSHRQSHQDEETGHGQRGSIKEMSHHPPGHTVVKDRSHLRKDDSEEEKEKEEDPGSHEEDDESSEQGEKGTHHGSRDQEDEEDEEEGHGLSLNQEEEEEEDKEEEEEEEDEERREERAEVGAPLSPDHSEEEEEEEEGLEEDEPRFTIIPNPLDRREEAGGASSEEESGEDTGPQDAQEYGNYQPGSLCGYCSFCNRCTECESCHCDEENMGEHCDQCQHCQFCYLCPLVCETVCAPGSYVDYFSSSLYQALADMLETPEP trdn_rabit Triadin TRDN protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MTEITAEGNASTTTTVIDSKNGSVPKSPGKVLKRTVTEDLVTTFSSPAAWLLVIALIITWSAVAVVMFDLVDYKNFSASSIAKMGSDPLKLVHDAVEETTDWIYGFFSLLSDIISSDGDEEDDEGDEDTAKGEIEEPPLKRKDIHKEKIEKQEKPERKIPTKVVHKEKEKEKEKVKEKEKPEKKATHKEKLEKKEKPETKTVTKEEKKARTKEKIEEKTKKEVKGVKQEKVKQTVAKAKEVQKTPKPKEKESKETAAVSKQEQKDQYAFCRYMIDIFVHGDLKPGQSPAIPPPSPTEQASRPTPALPTPEEKEGEKKKAEKKVTTETKKKAEKEDAKKKSEKETDIDMKKKEPGKSPDTKPGTVKVTTQAATKKDEKKEDSKKAKKPAEEQPKGKKQEKKEKHEEPAKSTKKEHAAPSEKQAKAKIERKEEVSAASTKKAVPAKKEEKTTKTVEQETRKEKPGKISSVLKDKELTKEKEVKVPASLKEKGSETKKDEKTSKPEPQIKKEEKPGKEVKPKPPQPQIKKEEKPEQDIMKPEKTALHGKPEEKVLKQVKAVTTEKHVKPKPAKKAEHQEKEPPSIKTDKPKSTSKGMPEVTESGKKKIEKSEKEIKVPARRESHQLQNVTKAEKPARGSKEGFEDVPASKKAKEEAEEVSSTKKQKSPISFFQCVYLDGYNGYGFQFPVTPAQYPGESSGKPNSPGPKQ mint May be involved in anchoring calsequestrin to the junctional sarcoplasmic reticulum and allowing its functional coupling with the ryanodine receptor trdn 1555537 n/a Triadin 1555539 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey n/a HRC 1555540 polyclonal western polyclonal antibody western blot polyclonal western unspecified role unspecified role prey prey cell lysate cell lysate P23327:p.Ser96Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false rate decreasing physical association physical association false false false cardiac homogenate Fig 6A and B p23327-trdn 1555538 n/a Triadin 1555541 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a HRC 1555540 polyclonal western polyclonal antibody western blot polyclonal western unspecified role unspecified role bait bait cell lysate cell lysate P23327:p.Ser96Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false rate decreasing region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 6C and D