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/ liu-2006a-1 Domain interaction sites of human lens betaB2-crystallin. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Liu BF., Liang JJ. BioCreative - Critical Assessment of Information Extraction systems in Biology A mammalian two hybrid approach was used and SEAP reporter gene assay activity was detected 48 h after transfection using the BD Great EscAPe SEAP fluorescence detection kit. The test protein (bait) was fused into the GAL4 DNA-BD in the pM vector, and the second test protein (prey) was fused into the VP16-AD in the pVP16 vector. The third vector, pG5SEAP, contains a reporter construct and is encoded with secreted alkaline phosphatase (SEAP),2 an enzyme that enables sampling of culture medium without cell lysis. jliang@rics.bwh.harvard.edu. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation liu-2006a-2 Domain interaction sites of human lens betaB2-crystallin. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Liu BF., Liang JJ. BioCreative - Critical Assessment of Information Extraction systems in Biology A mammalian two hybrid approach was used and SEAP reporter gene assay activity was detected 48 h after transfection using the BD Great EscAPe SEAP fluorescence detection kit. The test protein (bait) was fused into the GAL4 DNA-BD in the pM vector, and the second test protein (prey) was fused into the VP16-AD in the pVP16 vector. The third vector, pG5SEAP, contains a reporter construct and is encoded with secreted alkaline phosphatase (SEAP),2 an enzyme that enables sampling of culture medium without cell lysis. jliang@rics.bwh.harvard.edu. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation crbb2_human Beta-crystallin B2 Beta-B2 crystallin Beta-crystallin Bp CRYBB2 CRYB2 CRYB2A protein protein human Homo sapiens Human MASDHQTQAGKPQSLNPKIIIFEQENFQGHSHELNGPCPNLKETGVEKAGSVLVQAGPWVGYEQANCKGEQFVFEKGEYPRWDSWTSSRRTDSLSSLRPIKVDSQEHKIILYENPNFTGKKMEIIDDDVPSFHAHGYQEKVSSVRVQSGTWVGYQYPGYRGLQYLLEKGDYKDSSDFGAPHPQVQSVRRIRDMQWHQRGAFHPSN crybb2-crybb2 Mammalian 2 hybrid analysis reveals dimer formation by beta B2 Crystallin 499157 N/A 499159 unspecified role unspecified role prey prey P43320:p.Val54Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val60Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val73Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu97Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile109Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile125Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val144Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val152Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu166Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val187Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false N/A 499159 unspecified role unspecified role bait bait P43320:p.Val187Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val54Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val60Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val73Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu97Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile109Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile125Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val144Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu166Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 3 The authors used N-terminal and C-terminal regions in separate constructs to analyse dimer formation using a mammalian two hybrid assay. However the coordiates of the constructs are not given and the results are presented as if a full length beta B2 crysatllin protein was analysed. crybb2-crybb2-1 Mammalian 2 hybrid analysis reveals dimer formation by beta B2 Crystallin 499157 N/A 499159 unspecified role unspecified role bait bait N/A 499159 unspecified role unspecified role prey prey P43320:p.Val54Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val60Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val73Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu97Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile109Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile125Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val144Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val152Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu166Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val187Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 2 The authors used N-terminal and C-terminal regions in separate constructs to analyse dimer formation using a mammalian two hybrid assay. However the coordiates of the constructs are not given and the results are presented as if a full length beta B2 crysatllin protein was analysed. It is not clear form the text if bait or prey protein was muated in the analysis. The data is presented as if the bait protein was mutated. crybb2-crybb2-2 Mammalian 2 hybrid analysis reveals dimer formation by beta B2 Crystallin 499157 N/A 499159 unspecified role unspecified role bait bait P43320:p.Val54Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val60Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val73Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu97Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile109Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile125Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val144Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu166Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val187Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false N/A 499159 unspecified role unspecified role prey prey P43320:p.Val54Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val60Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val73Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu97Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile109Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Ile125Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val144Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val152Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Leu166Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val187Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 1 The authors used N-terminal and C-terminal regions in separate constructs to analyse dimer formation using a mammalian two hybrid assay. However the coordiates of the constructs are not given and the results are presented as if a full length beta B2 crysatllin protein was analysed. crybb2-crybb2-3 Size exclusion analysis reveals dimer formation by beta B2 Crystallin 499158 N/A 499159 unspecified role unspecified role prey prey P43320:p.Val144Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val60Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false N/A 499159 unspecified role unspecified role bait bait P43320:p.Val60Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P43320:p.Val144Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 5