bhf-ucl Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. J. Biol. Chem. (0021-9258) 2008 imex curation Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting sequence tag sequence tag identification sequence tag Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. COS-1 cell lysates were separated by a first horizontal dimension using a wide pH range isoelectrofocusing gel strip and then vertically by SDS-PAGE. After the SDS-PAGE, the proteins were electrotransferred onto a nitrocellulose membrane and probed by Far Western with anti-V5 antibodies. seidahn@ircm.qc.ca Accepted 2014-JUL-02 by MGT J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. J. Biol. Chem. (0021-9258) 2008 imex curation Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca Accepted 2014-JUL-02 by MGT J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. J. Biol. Chem. (0021-9258) 2008 imex curation Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting weight by staining molecular weight estimation by staining weight by staining Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins The COS -1 line in fig. 1B is a control that shows no interaction when PCSK9 is not present in the conditioned medium (pIRES-V5) After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca Accepted 2014-JUL-02 by MGT J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. J. Biol. Chem. (0021-9258) 2008 imex curation Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip ms participant Identification by mass spectrometry ms participant Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca Accepted 2014-JUL-02 by MGT J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. J. Biol. Chem. (0021-9258) 2008 imex curation Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro electrophoresis electrophoretic mobility-based method electrophoresis western blot western blot Immuno blot Immuno blot Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca Accepted 2014-JUL-02 by MGT J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. J. Biol. Chem. (0021-9258) 2008 imex curation Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 confocal microscopy confocal microscopy anti-tag immunocytochemistry anti-tag immunocytochemistry Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca Accepted 2014-JUL-02 by MGT J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. Annexin A2 is a C-terminal PCSK9-binding protein that regulates endogenous low density lipoprotein receptor levels. J. Biol. Chem. (0021-9258) 2008 imex curation Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. seidahn@ircm.qc.ca 21-Jul-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Mayer G., Poirier S., Seidah N.G. Cardiac - Interactions involving cardiac related proteins After talking with Sandra she agreed to annotate the experiments in fig. 3A, 4B and 9. AnxA2-His6 was confirmed to be human through correspondence with the authors and by looking at the EMD bioscience catalog where they bought it from. seidahn@ircm.qc.ca Accepted 2014-JUL-02 by MGT J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation anxa2_human Annexin A2 ANXA2 ANX2 ANX2L4 CAL1H LPC2D Annexin-2 Annexin II Lipocortin II Calpactin I heavy chain Chromobindin-8 p36 Protein I Placental anticoagulant protein IV Calpactin-1 heavy chain protein protein human Homo sapiens Human MSTVHEILCKLSLEGDHSTPPSAYGSVKAYTNFDAERDALNIETAIKTKGVDEVTIVNILTNRSNAQRQDIAFAYQRRTKKELASALKSALSGHLETVILGLLKTPAQYDASELKASMKGLGTDEDSLIEIICSRTNQELQEINRVYKEMYKTDLEKDIISDTSGDFRKLMVALAKGRRAEDGSVIDYELIDQDARDLYDAGVKRKGTDVPKWISIMTERSVPHLQKVFDRYKSYSPYDMLESIRKEVKGDLENAFLNLVQCIQNKPLYFADRLYDSMKGKGTRDKVLIRIMVSRSEVDMLKIRSEFKRKYGKSLYYYIQQDTKGDYQKALLYLCGGDD 5126E1337A0CBEA1 pcsk9_human Proprotein convertase subtilisin/kexin type 9 Subtilisin/kexin-like protease PC9 Neural apoptosis-regulated convertase 1 Proprotein convertase 9 PCSK9 NARC1 PSEC0052 protein protein human Homo sapiens Human MGTVSSRRSWWPLPLLLLLLLLLGPAGARAQEDEDGDYEELVLALRSEEDGLAEAPEHGTTATFHRCAKDPWRLPGTYVVVLKEETHLSQSERTARRLQAQAARRGYLTKILHVFHGLLPGFLVKMSGDLLELALKLPHVDYIEEDSSVFAQSIPWNLERITPPRYRADEYQPPDGGSLVEVYLLDTSIQSDHREIEGRVMVTDFENVPEEDGTRFHRQASKCDSHGTHLAGVVSGRDAGVAKGASMRSLRVLNCQGKGTVSGTLIGLEFIRKSQLVQPVGPLVVLLPLAGGYSRVLNAACQRLARAGVVLVTAAGNFRDDACLYSPASAPEVITVGATNAQDQPVTLGTLGTNFGRCVDLFAPGEDIIGASSDCSTCFVSQSGTSQAAAHVAGIAAMMLSAEPELTLAELRQRLIHFSAKDVINEAWFPEDQRVLTPNLVAALPPSTHGAGWQLFCRTVWSAHSGPTRMATAVARCAPDEELLSCSSFSRSGKRRGERMEAQGGKLVCRAHNAFGGEGVYAIARCCLLPQANCSVHTAPPAEASMGTRVHCHQQGHVLTGCSSHWEVEDLGTHKPPVLRPRGQPNQCVGHREASIHASCCHAPGLECKVKEHGIPAPQEQVTVACEEGWTLTGCSALPGTSHVLGAYAVDNTCVVRSRDVSTTGSTSEGAVTAVAICCRSRHLAQASQELQ mint homomint May be implicated in the differentiation of cortical neurons and may play a role in cholesterol homeostasis Defects in PCSK9 are the cause of familial hypercholesterolemia 3 (FH3) [MIM:603776]. FH3 inheritance is autosomal dominant 9BCB9418B90AEE23 anxa2_chlae_protein Annexin A2 protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey similar to P07355 pcsk9-anxa2 3 9 unspecified role unspecified role bait bait conditioned medium conditioned medium c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q8NBP7:p.Gln554Glu mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain Q E Wed May 02 15:58:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln554glu' Tue Jun 30 18:55:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln554Glu' crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 8 unspecified role unspecified role prey prey cell lysate cell lysate crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 physical association physical association Fig. 3A, 4B anxa2-pcsk9-1 7 9 unspecified role unspecified role prey prey conditioned medium conditioned medium c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 8 unspecified role unspecified role bait bait purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fg. 3B pcsk9-anxa2-1 2 9 unspecified role unspecified role bait bait c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role prey prey c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 3C pcsk9-anxa2chlae-1 4 9 unspecified role unspecified role bait bait c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 10 unspecified role unspecified role prey prey physical association physical association Fig. 2 A,E pcsk9-anxa2-2 3 9 unspecified role unspecified role bait bait conditioned medium conditioned medium c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 8 unspecified role unspecified role prey prey purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 3D Purified AnxA2 was submitted to electrophoresis under reducing and non-reducing conditions. PCSK9 binds in a dose-dependent manner AnxA2 in both situations. Under reducing conditions only binding of the monomeric form of Anxa-His to Pcsk9 was observed. anxa2-anxa2 5 8 unspecified role unspecified role unspecified role unspecified role purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 3D reducing conditions pcsk9-anxa2chlae-3 1 9 unspecified role unspecified role bait bait conditioned medium conditioned medium c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 10 unspecified role unspecified role prey prey cell lysate cell lysate chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 physical association physical association Fig. 2C, D, E anxa2-pcsk9-2 6 9 unspecified role unspecified role unspecified role unspecified role c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role unspecified role unspecified role c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Fig. 5 pcsk9-anxa2-6 3 9 unspecified role unspecified role bait bait conditioned medium conditioned medium c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 8 unspecified role unspecified role prey prey cell lysate cell lysate c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range r1 domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain P07355:p.[Arg77Ala;Arg78Ala;Lys80Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain R A Fri Jul 22 11:05:46 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg78ala,lys80ala,arg77ala' Wed May 02 19:16:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg77ala,arg78ala,lys80ala' Tue Jun 30 18:41:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg77Ala;Arg78Ala;Lys80Ala]' P07355:p.[Arg77Ala;Arg78Ala;Lys80Ala;Lys81Ala;Lys88Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Fri Jul 22 10:49:36 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys88ala,arg77ala,arg78ala,lys80ala,lys81ala' Wed May 02 16:37:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg77ala,arg78ala,lys80ala,lys81ala,lys88ala' Tue Jun 30 20:19:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg77Ala;Arg78Ala;Lys80Ala;Lys81Ala;Lys88Ala]' P07355:p.[Arg77Ala;Arg78Ala;Lys80Ala;Lys81Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 14:31:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg77ala,arg78ala,lys80ala,lys81ala' Tue Jun 30 18:22:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg77Ala;Arg78Ala;Lys80Ala;Lys81Ala]' 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. physical association physical association Fig. 9 B, C Individual deletions of R2, R3 and R4 reduce the ability to bind to PCSK9 Exchanging the 49-75aa fragment in AnxA2 with the 58-84aa fragment of AnxA1 significantly reduces the AnxA2 ability to bind PCSK9. (Fig. 9C) pcsk9-anxa2-7 3 9 unspecified role unspecified role bait bait conditioned medium conditioned medium c-terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 8 unspecified role unspecified role prey prey purified purified n-terminus s tag s tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 3D Purified AnxA2 was submitted to electrophoresis under reducing and non-reducing conditions. PCSK9 binds in a dose-dependent manner AnxA2 in both situations.