UniProt UniProt http://www.uniprot.org EFHB is a Novel Cytosolic Ca2+ Sensor That Modulates STIM1-SARAF Interaction. EFHB is a Novel Cytosolic Ca2+ Sensor That Modulates STIM1-SARAF Interaction. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 2017 imex curation Albarran L., Lopez JJ., Jardin I., Sanchez-Collado J., Berna-Erro A., Smani T., Camello PJ., Salido GM., Rosado JA. Source of proteins taken from previous work of the same authors (SARAF PMID: 27068144 and STIM1 PMID: 30975919) also PMID: 23943619· ijp@unex.es Only protein-protein interactions imex curation 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 western blot western blot Immuno blot Immuno blot Albarran L., Lopez JJ., Jardin I., Sanchez-Collado J., Berna-Erro A., Smani T., Camello PJ., Salido GM., Rosado JA. Source of proteins taken from previous work of the same authors (SARAF PMID: 27068144 and STIM1 PMID: 30975919) also PMID: 23943619· ijp@unex.es Accepted 2021-JUL-20 AT 13:40 BST AT 13:40 BST by MGT Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 2017 Only protein-protein interactions imex curation imex curation EFHB is a Novel Cytosolic Ca2+ Sensor That Modulates STIM1-SARAF Interaction. EFHB is a Novel Cytosolic Ca2+ Sensor That Modulates STIM1-SARAF Interaction. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 2017 imex curation Albarran L., Lopez JJ., Jardin I., Sanchez-Collado J., Berna-Erro A., Smani T., Camello PJ., Salido GM., Rosado JA. Source of proteins taken from previous work of the same authors (SARAF PMID: 27068144 and STIM1 PMID: 30975919) also PMID: 23943619· ijp@unex.es Only protein-protein interactions imex curation human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Albarran L., Lopez JJ., Jardin I., Sanchez-Collado J., Berna-Erro A., Smani T., Camello PJ., Salido GM., Rosado JA. Source of proteins taken from previous work of the same authors (SARAF PMID: 27068144 and STIM1 PMID: 30975919) also PMID: 23943619· ijp@unex.es Accepted 2021-JUL-20 AT 13:40 BST AT 13:40 BST by MGT Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 2017 Only protein-protein interactions imex curation imex curation saraf_human Store-operated calcium entry-associated regulatory factor SARAF HBV X-transactivated gene 3 protein HBV XAg-transactivated protein 3 Protein FOAP-7 TMEM66 Transmembrane protein 66 XTP3 HSPC035 NPD003 PSEC0019 UNQ1967/PRO4499 protein protein human Homo sapiens Human MAAACGPGAAGYCLLLGLHLFLLTAGPALGWNDPDRMLLRDVKALTLHYDRYTTSRRLDPIPQLKCVGGTAGCDSYTPKVIQCQNKGWDGYDVQWECKTDLDIAYKFGKTVVSCEGYESSEDQYVLRGSCGLEYNLDYTELGLQKLKESGKQHGFASFSDYYYKWSSADSCNMSGLITIVVLLGIAFVVYKLFLSDGQYSPPPYSEYPPFSHRYQRFTNSAGPPPPGFKSEFTGPQNTGHGATSGFGSAFTGQQGYENSGPGFWTGLGTGGILGYLFGSNRAATPFSDSWYYPSYPPSYPGTWNRAYSPLHGGSGSYSVCSNSDTKTRTASGYGGTRRR 83CC0517AB635FE9 stim1_human Stromal interaction molecule 1 STIM1 GOK protein protein human Homo sapiens Human MDVCVRLALWLLWGLLLHQGQSLSHSHSEKATGTSSGANSEESTAAEFCRIDKPLCHSEDEKLSFEAVRNIHKLMDDDANGDVDVEESDEFLREDLNYHDPTVKHSTFHGEDKLISVEDLWKAWKSSEVYNWTVDEVVQWLITYVELPQYEETFRKLQLSGHAMPRLAVTNTTMTGTVLKMTDRSHRQKLQLKALDTVLFGPPLLTRHNHLKDFMLVVSIVIGVGGCWFAYIQNRYSKEHMKKMMKDLEGLHRAEQSLHDLQERLHKAQEEHRTVEVEKVHLEKKLRDEINLAKQEAQRLKELREGTENERSRQKYAEEELEQVREALRKAEKELESHSSWYAPEALQKWLQLTHEVEVQYYNIKKQNAEKQLLVAKEGAEKIKKKRNTLFGTFHVAHSSSLDDVDHKILTAKQALSEVTAALRERLHRWQQIEILCGFQIVNNPGIHSLVAALNIDPSWMGSTRPNPAHFIMTDDVDDMDEEIVSPLSMQSPSLQSSVRQRLTEPQHGLGSQRDLTHSDSESSLHMSDRQRVAPKPPQMSRAADEALNAMTSNGSHRLIEGVHPGSLVEKLPDSPALAKKALLALNHGLDKAHSLMELSPSAPPGGSPHLDSSRSHSPSSPDPDTPSPVGDSRALQASRNTRIPHLAGKKAVAEEDNGSIGEETDSSPGRKKFPLKIFKKPLKK DIP protein Q13586 original sequence version: 141 DIP protein Q13586 original sequence version: 99 DIP protein Q13586 original sequence version: 108 DIP protein Q13586 original sequence version: 121 DIP protein Q13586 original sequence version: 125 DIP protein Q13586 original sequence version: 100 FFB5E7CB3D68A7A6 efhb_human EF-hand domain-containing family member B EFHB protein protein human Homo sapiens Human MNMEIGHPHEGKDDLGDKRVIMGTKFPMELGIRVGLGKEDSRCGESPVVSNKCEGRMAPPETKFPLSKGLEMGLERQNISRTVMQRGSLGVDSVSASQGTKPSLLPGRMGLENESLLAGYTHERIIQPPLGRVCGSSQAAGSRRAPLASGPEGVEELVGKPAFVMEPRQEMEKESTCVLMKPNTEIKLPVEVDIGLTQAEGPDETKNTEPQMGLVIEPPQCQFAQQHEQRKEAGNIESGVEPPDRIRPIYSGKFFDRTPCWPSAGKVIPVGYRVATCLTEKLPRLITPPEAKKYFNFRYPPAGVERVFYGRANDPQIAPYLTHGIRSKISVLANTLINPQPITTFQQKIKDKKESIYLSNRRAPLGKSHDQAPGLPKGMDTTNTTFGTAVIKEYSAKDVVNPPKSYEEVFKEGNEGHDLYVVSHNDYYAGEAKNRKYNPSSFHRCSVYGVPTPHFNDGRAMAKSLYWLHELQMKRGAKFVSKRADDFKEKFQHKLGRVLDPIAETMNVPPDCTFGACLRPEEYGVGDLIHNRLPDEYLRGKDRQRALIAAVRHHLKKVNYQKFDTLLAAFRHYDKKGDGMIDKDELQEACDQANLSLDDKLLDQLFDYCDVDNDGFINYLEFANFLNWKDKMLLKEYEERVIIKGRKPDCVNPTEANVEEPEQTLLIKPEDIVLKEAGSTEKTLRTLLRPSDKVSNYYKTTSSEINAIVGAIPSTCYPICGVPTIRSDIPAPRIRRISDRTNYGEEGSAYSLLYPTIFARKGVFERDFFKTRSKEEIAEILCNIGVKLSDEEFENVWNLASKKHHRGEVCVENIRNVLDELRHADRIKCKTLM crcm1_human Calcium release-activated calcium channel protein 1 ORAI1 CRACM1 TMEM142A Protein orai-1 Transmembrane protein 142A protein protein human Homo sapiens Human MHPEPAPPPSRSSPELPPSGGSTTSGSRRSRRRSGDGEPPGAPPPPPSAVTYPDWIGQSYSEVMSLNEHSMQALSWRKLYLSRAKLKASSRTSALLSGFAMVAMVEVQLDADHDYPPGLLIAFSACTTVLVAVHLFALMISTCILPNIEAVSNVHNLNSVKESPHERMHRHIELAWAFSTVIGTLLFLAEVVLLCWVKFLPLKKQPGQPRPTSKPPASGAAANVSTSGITPGQAAAIASTTIMVPFGLIFIVFAVHFYRSLVSHKTDRQFQELNELAEFARLQDQLDHRGDHPLTPGSHYA DIP protein Q96D31 original sequence version: 95 DIP protein Q96D31 original sequence version: 38 DIP protein Q96D31 original sequence version: 58 DIP protein Q96D31 original sequence version: 55 DIP protein Q96D31 original sequence version: 57 DIP protein Q96D31 original sequence version: 53 DIP protein Q96D31 original sequence version: 63 DIP protein Q96D31 original sequence version: 76 DIP protein Q96D31 original sequence version: 79 DIP protein Q96D31 original sequence version: 56 F85F7C06F36628D4 orai1-efhb-1 1 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate physical association physical association Fig.2A Thapsigargin (CHEBI:9516) stim1-efhb-1 1 5 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate physical association physical association Fig.2A Thapsigargin (CHEBI:9516) stim1-orai1-2 1 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate physical association physical association Fig.3A, 4B Thapsigargin (CHEBI:9516) Effect of Thapsigargin was overcome by depletion of EFHB The interaction was increased in cells expressing EFHB (residues 165-833) stim1-orai1-3 2 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate physical association physical association Fig.3B Thapsigargin (CHEBI:9516) Effect of Thapsigargin was overcome by depletion of EFHB and increased in cells expressing EFHB (residues 165-833) orai1-efhb-2 2 5 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 6 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig.6C Thapsigargin (CHEBI:9516) dimethyl BAPTA impaired the TG-evoked increase in the EFHB-Orai1 association. stim1-efhb-2 2 3 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level association association Fig.6B Treatment of the cells with Thapsigargin (CHEBI:9516) resulted in a significant initial decrease in SARAF-STIM1 interaction. Pretreatment of the cells with dimethy-BAPTA (CHEBI:60889) impaired the recovery of the SARAF-STIM1 interaction after 30 minutes. stim1-saraf-1 2 3 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig.7A The inhibitory effect of Thapsigargin on this interaction was lost in EFHB depleted cells Thapsigargin (CHEBI:9516) stim1-efhb-3 2 5 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role bait bait cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig.6D Overexpression of SARAF significantly attenuated the TG-induced EFHB-STIM1 interaction.