MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Structure of the Ire1 autophosphorylation complex and implications for the unfolded protein response. Structure of the Ire1 autophosphorylation complex and implications for the unfolded protein response. EMBO J. (0261-4189) 2011 imex curation Ali MM., Bagratuni T., Davenport EL., Nowak PR., Silva-Santisteban MC., Hardcastle A., McAndrews C., Rowlands MG., Morgan GJ., Aherne W., Collins I., Davies FE., Pearl LH. maruf.ali@imperial.ac.uk Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Ali MM., Bagratuni T., Davenport EL., Nowak PR., Silva-Santisteban MC., Hardcastle A., McAndrews C., Rowlands MG., Morgan GJ., Aherne W., Collins I., Davies FE., Pearl LH. maruf.ali@imperial.ac.uk EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Structure of the Ire1 autophosphorylation complex and implications for the unfolded protein response. Structure of the Ire1 autophosphorylation complex and implications for the unfolded protein response. EMBO J. (0261-4189) 2011 imex curation Ali MM., Bagratuni T., Davenport EL., Nowak PR., Silva-Santisteban MC., Hardcastle A., McAndrews C., Rowlands MG., Morgan GJ., Aherne W., Collins I., Davies FE., Pearl LH. maruf.ali@imperial.ac.uk Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Ali MM., Bagratuni T., Davenport EL., Nowak PR., Silva-Santisteban MC., Hardcastle A., McAndrews C., Rowlands MG., Morgan GJ., Aherne W., Collins I., Davies FE., Pearl LH. maruf.ali@imperial.ac.uk EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation ern1_human Serine/threonine-protein kinase/endoribonuclease IRE1 ERN1 Inositol-requiring protein 1 Ire1-alpha Endoplasmic reticulum-to-nucleus signaling 1 IRE1 protein protein human Homo sapiens Human MPARRLLLLLTLLLPGLGIFGSTSTVTLPETLLFVSTLDGSLHAVSKRTGSIKWTLKEDPVLQVPTHVEEPAFLPDPNDGSLYTLGSKNNEGLTKLPFTIPELVQASPCRSSDGILYMGKKQDIWYVIDLLTGEKQQTLSSAFADSLCPSTSLLYLGRTEYTITMYDTKTRELRWNATYFDYAASLPEDDVDYKMSHFVSNGDGLVVTVDSESGDVLWIQNYASPVVAFYVWQREGLRKVMHINVAVETLRYLTFMSGEVGRITKWKYPFPKETEAKSKLTPTLYVGKYSTSLYASPSMVHEGVAVVPRGSTLPLLEGPQTDGVTIGDKGECVITPSTDVKFDPGLKSKNKLNYLRNYWLLIGHHETPLSASTKMLERFPNNLPKHRENVIPADSEKKSFEEVINLVDQTSENAPTTVSRDVEEKPAHAPARPEAPVDSMLKDMATIILSTFLLIGWVAFIITYPLSMHQQQQLQHQQFQKELEKIQLLQQQQQQLPFHPPGDTAQDGELLDTSGPYSESSGTSSPSTSPRASNHSLCSGSSASKAGSSPSLEQDDGDEETSVVIVGKISFCPKDVLGHGAEGTIVYRGMFDNRDVAVKRILPECFSFADREVQLLRESDEHPNVIRYFCTEKDRQFQYIAIELCAATLQEYVEQKDFAHLGLEPITLLQQTTSGLAHLHSLNIVHRDLKPHNILISMPNAHGKIKAMISDFGLCKKLAVGRHSFSRRSGVPGTEGWIAPEMLSEDCKENPTYTVDIFSAGCVFYYVISEGSHPFGKSLQRQANILLGACSLDCLHPEKHEDVIARELIEKMIAMDPQKRPSAKHVLKHPFFWSLEKQLQFFQDVSDRIEKESLDGPIVKQLERGGRAVVKMDWRENITVPLQTDLRKFRTYKGGSVRDLLRAMRNKKHHYRELPAEVRETLGSLPDDFVCYFTSRFPHLLAHTYRAMELCSHERLFQPYYFHEPPEPQPPVTPDAL A2DF808CCE015536 s2 RRRLSFAEPG peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) RRRLSFAEPG mint 75BAF11DD9D5B734 ern1-1 1 Ire1 alpha 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 2.0 direct interaction direct interaction mint The arrangement of molecules in the human Ire1α crystals is quite different to that of the yeast Ire1 structures. Only two protein protein interfaces of any substance are observed. The largest (burying >1700 Å2 of molecular surface) is an approximately two-fold symmetric parallel interaction, involving residues from the kinase domain only, and generates a face-to-face dimer arrangement in which the open mouths of the interlobe clefts, containing bound Mg2+-ADP, face each other across the dimer (Figure 2A). Coordinates and structure factors have been deposited in the Protein Databank with accession code: 3P23 f1 f2a t1 ern1-1 2 Ire1 alpha 3 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined self self self self purified purified phosphorylated phosphorylated residue phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain resulting-ptm phosphorylation phosphorylation reaction phosphorylation true mint f2b f4a f4b true ern1-s2 2 S2 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined enzyme target enzyme target substrate substrate neutral component neutral component purified purified biotin tag biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Ire1 alpha 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified phosphorylation phosphorylation reaction phosphorylation f4e mint To date, no other biological substrate for Ire1α phosphorylation other than itself, has been identified. However, using a commercial homogeneous time-resolved fluorescence assay system (Cisbio, France), we identified a peptide, S2, that was efficiently phosphorylated by Ire1α (Figure 4E).