IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ shiroishi-2006-6 in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Shiroishi M., Kuroki K., Ose T., Rasubala L., Shiratori I., Arase H., Tsumoto K., Kumagai I., Kohda D., Maenaka K. Details of B2M and HLA-G constructs have been taken from PDB entry 2d31. kmaenaka@bioreg.kyushu-u.ac.jp The Journal of biological chemistry 2006 imex curation Only protein-protein interactions imex curation shiroishi-2006-1 in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Shiroishi M., Kuroki K., Ose T., Rasubala L., Shiratori I., Arase H., Tsumoto K., Kumagai I., Kohda D., Maenaka K. Details of B2M and HLA-G constructs have been taken from PDB entry 2d31. kmaenaka@bioreg.kyushu-u.ac.jp The Journal of biological chemistry 2006 imex curation Only protein-protein interactions imex curation shiroishi-2006-4 in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight by staining molecular weight estimation by staining weight by staining Shiroishi M., Kuroki K., Ose T., Rasubala L., Shiratori I., Arase H., Tsumoto K., Kumagai I., Kohda D., Maenaka K. Details of B2M and HLA-G constructs have been taken from PDB entry 2d31. kmaenaka@bioreg.kyushu-u.ac.jp The Journal of biological chemistry 2006 imex curation Only protein-protein interactions imex curation shiroishi-2006-2 ecoli Escherichia coli (strain K12) comigration in sds comigration in sds page comigration in sds predetermined predetermined participant predetermined Shiroishi M., Kuroki K., Ose T., Rasubala L., Shiratori I., Arase H., Tsumoto K., Kumagai I., Kohda D., Maenaka K. SDS-PAGE of the HLA-G under nonreducing conditions. Details of B2M and HLA-G constructs have been taken from PDB entry 2d31. kmaenaka@bioreg.kyushu-u.ac.jp The Journal of biological chemistry 2006 imex curation Only protein-protein interactions imex curation shiroishi-2006-3 ecoli Escherichia coli (strain K12) comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight by staining molecular weight estimation by staining weight by staining Shiroishi M., Kuroki K., Ose T., Rasubala L., Shiratori I., Arase H., Tsumoto K., Kumagai I., Kohda D., Maenaka K. Details of B2M and HLA-G constructs have been taken from PDB entry 2d31. kmaenaka@bioreg.kyushu-u.ac.jp The Journal of biological chemistry 2006 imex curation Only protein-protein interactions imex curation shiroishi-2006-5 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Shiroishi M., Kuroki K., Ose T., Rasubala L., Shiratori I., Arase H., Tsumoto K., Kumagai I., Kohda D., Maenaka K. Details of B2M and HLA-G constructs have been taken from PDB entry 2d31. kmaenaka@bioreg.kyushu-u.ac.jp The Journal of biological chemistry 2006 imex curation Only protein-protein interactions imex curation h2ax_human Histone H2AX H2AX H2AFX Histone H2A.X protein protein human Homo sapiens Human MSGRGKTGGKARAKAKSRSSRAGLQFPVGRVHRLLRKGHYAERVGAGAPVYLAAVLEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRNDEELNKLLGGVTIAQGGVLPNIQAVLLPKKTSATVGPKAPSGGKKATQASQEY lirb1_human Leukocyte immunoglobulin-like receptor subfamily B member 1 ILT2 LIR1 MIR7 CD85 antigen-like family member J Immunoglobulin-like transcript 2 Monocyte/macrophage immunoglobulin-like receptor 7 LILRB1 protein protein human Homo sapiens Human MTPILTVLICLGLSLGPRTHVQAGHLPKPTLWAEPGSVITQGSPVTLRCQGGQETQEYRLYREKKTALWITRIPQELVKKGQFPIPSITWEHAGRYRCYYGSDTAGRSESSDPLELVVTGAYIKPTLSAQPSPVVNSGGNVILQCDSQVAFDGFSLCKEGEDEHPQCLNSQPHARGSSRAIFSVGPVSPSRRWWYRCYAYDSNSPYEWSLPSDLLELLVLGVSKKPSLSVQPGPIVAPEETLTLQCGSDAGYNRFVLYKDGERDFLQLAGAQPQAGLSQANFTLGPVSRSYGGQYRCYGAHNLSSEWSAPSDPLDILIAGQFYDRVSLSVQPGPTVASGENVTLLCQSQGWMQTFLLTKEGAADDPWRLRSTYQSQKYQAEFPMGPVTSAHAGTYRCYGSQSSKPYLLTHPSDPLELVVSGPSGGPSSPTTGPTSTSGPEDQPLTPTGSDPQSGLGRHLGVVIGILVAVILLLLLLLLLFLILRHRRQGKHWTSTQRKADFQHPAGAVGPEPTDRGLQWRSSPAADAQEENLYAAVKHTQPEDGVEMDTRSPHDEDPQAVTYAEVKHSRPRREMASPPSPLSGEFLDTKDRQAEEDRQMDTEAAASEAPQDVTYAQLHSLTLRREATEPPPSQEGPSPAVPSIYATLAIH hlag_human HLA class I histocompatibility antigen, alpha chain G HLAG HLA-G HLA-6.0 HLA G antigen MHC class I antigen G protein protein human Homo sapiens Human MVVMAPRTLFLLLSGALTLTETWAGSHSMRYFSAAVSRPGRGEPRFIAMGYVDDTQFVRFDSDSACPRMEPRAPWVEQEGPEYWEEETRNTKAHAQTDRMNLQTLRGYYNQSEASSHTLQWMIGCDLGSDGRLLRGYEQYAYDGKDYLALNEDLRSWTAADTAAQISKRKCEAANVAEQRRAYLEGTCVEWLHRYLENGKEMLQRADPPKTHVTHHPVFDYEATLRCWALGFYPAEIILTWQRDGEDQTQDVELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPEPLMLRWKQSSLPTIPIMGIVAGLVVLAAVVTGAAVAAVLWRKKSSD p61769-pro_0000018779 Beta-2-microglobulin B2M CDABP0092 HDCMA22P protein protein human Homo sapiens Human IQRTPKIQVYSRHPAENGKSNFLNCYVSGFHPSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEYACRVNHVTLSQPKIVKWDRDM 21 119 lirb2_human Leukocyte immunoglobulin-like receptor subfamily B member 2 LILRB2 ILT4 LIR2 MIR10 CD85 antigen-like family member D Immunoglobulin-like transcript 4 Monocyte/macrophage immunoglobulin-like receptor 10 protein protein human Homo sapiens Human MTPIVTVLICLGLSLGPRTHVQTGTIPKPTLWAEPDSVITQGSPVTLSCQGSLEAQEYRLYREKKSASWITRIRPELVKNGQFHIPSITWEHTGRYGCQYYSRARWSELSDPLVLVMTGAYPKPTLSAQPSPVVTSGGRVTLQCESQVAFGGFILCKEGEEEHPQCLNSQPHARGSSRAIFSVGPVSPNRRWSHRCYGYDLNSPYVWSSPSDLLELLVPGVSKKPSLSVQPGPVVAPGESLTLQCVSDVGYDRFVLYKEGERDLRQLPGRQPQAGLSQANFTLGPVSRSYGGQYRCYGAHNLSSECSAPSDPLDILITGQIRGTPFISVQPGPTVASGENVTLLCQSWRQFHTFLLTKAGAADAPLRLRSIHEYPKYQAEFPMSPVTSAHAGTYRCYGSLNSDPYLLSHPSEPLELVVSGPSMGSSPPPTGPISTPAGPEDQPLTPTGSDPQSGLGRHLGVVIGILVAVVLLLLLLLLLFLILRHRRQGKHWTSTQRKADFQHPAGAVGPEPTDRGLQWRSSPAADAQEENLYAAVKDTQPEDGVEMDTRAAASEAPQDVTYAQLHSLTLRRKATEPPPSQEREPPAEPSIYATLAIH b2m-h2afx-2 2973419 N/A 2973425 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 2973426 unspecified role unspecified role bait bait c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973427 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973428 unspecified role unspecified role prey prey whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 2973437 2973432 2973439 2973432 physical association physical association false false false 2973419 2 (A)B/E lilrb1-b2m-1 2973419 N/A 2973425 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 N/A 2973426 unspecified role unspecified role bait bait c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973428 unspecified role unspecified role prey prey whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 2973447 2973442 2973449 2973442 physical association physical association false false false 2 A The simple 1:1 (Langmuir) binding model was not successful at high concentrations of the dimer, but was well fitted with a bivalent analyte model. (1:2 (dimer:LILRB) complex ) affinity constant -Kd ∼6.7 nm for LILRB1, at low concentrations of dimer lilrb1-b2m-2 2973419 N/A 2973425 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 N/A 2973429 unspecified role unspecified role bait bait c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973428 unspecified role unspecified role prey prey whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 2973457 2973452 2973459 2973452 physical association physical association false false false 2 C The simple 1:1 (Langmuir) binding model was not successful at high concentrations of the dimer, but was well fitted with a bivalent analyte model. (1:2 (dimer:LILRB) complex ) affinity constant -Kd ∼750 nm for LILRB2, at low concentrations of dimer lilrb2-b2m-1 2973419 N/A 2973425 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 2973429 unspecified role unspecified role bait bait c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973427 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973428 unspecified role unspecified role prey prey whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 2973467 2973462 2973469 2973462 physical association physical association false false false 2973419 2 (C)/D/E lilrb1-b2m-3 2973419 N/A 2973425 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 N/A 2973426 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role bait bait c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973428 unspecified role unspecified role bait bait whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 2973477 2973472 2973479 2973472 physical association physical association false false false 2973419 Supp fig 1, Supp. table 1 lilrb1-b2m-4 2973419 N/A 2973425 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 2973426 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973427 unspecified role unspecified role bait bait ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973428 unspecified role unspecified role bait bait whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 2973486 2973482 2973489 2973482 physical association physical association false false false 2973419 Supp fig 1, Supp Table 1 lilrb2-b2m-2 2973419 N/A 2973425 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 N/A 2973429 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role bait bait c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973428 unspecified role unspecified role bait bait whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 2973497 2973492 2973499 2973492 physical association physical association false false false 2973419 Supp fig 1, Supp. table 1 lilrb2-b2m-3 2973419 N/A 2973425 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 2973429 unspecified role unspecified role prey prey ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973427 unspecified role unspecified role bait bait ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c_terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P17693:p.Cys66Ser mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 2973428 unspecified role unspecified role bait bait whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 2973506 2973502 2973510 2973502 physical association physical association false false false 2973419 Supp fig 1, Supp Table 1 hla_g-4 2973420 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P17693:p.Cys66Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 physical association physical association false false false 1A hla_g-lilrb1-1 2973420 N/A 2973426 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 physical association physical association false false false 1E The 1(dimer):2 (LILRB1) molar ratio sample showed only one complex peak and thus represented the true equilibrium complex stoichiometry. hla_g-lilrb2-3 2973420 N/A 2973429 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 physical association physical association false false false 1G The 1(dimer):2 (LILRB2) molar ratio sample showed only one complex peak and thus represented the true equilibrium complex stoichiometry. hla_g-7 2973421 N/A 2973426 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 physical association physical association false false false 1D the full complex was composed of one HLA-G dimer and two LILRB1s (1:2 complex). hla_g-lilrb1-2 2973421 N/A 2973429 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 physical association physical association false false false 1F the full complex was composed of one HLA-G dimer and two LILRB2s (1:2 complex). hla_g-5 2973422 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P17693:p.Cys66Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 physical association physical association false false false 1B Beta-2-microglobulin was also present on the gel. hla_g-6 2973423 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P17693:p.Cys66Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 physical association physical association false false false 1C b2m-h2afx-1 2973424 N/A 2973425 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 4.0 N/A 2973427 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 N/A 2973428 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 physical association physical association false false false Table 1 3.2 A RFree = 0.298 RWork = 0.235