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/ mast-2013-2 in vitro In vitro uv-vis ultraviolet-visible spectroscopy uv-vis predetermined predetermined participant predetermined Mast N., Zheng W., Stout CD., Pikuleva IA. dave@scripps.edu,iap8@case.edu The Journal of biological chemistry 2013 Only protein-protein interactions imex curation imex curation mast-2013-1 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Mast N., Zheng W., Stout CD., Pikuleva IA. dave@scripps.edu,iap8@case.edu The Journal of biological chemistry 2013 Only protein-protein interactions imex curation imex curation bicalutamide Bicalutamide, an androgen antagonist small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cp46a_human Cholesterol 24-hydroxylase CYP46A1 CYP46 Cytochrome P450 46A1 Cholesterol 24-monooxygenase Cholesterol 24S-hydroxylase protein protein human Homo sapiens Human MSPGLLLLGSAVLLAFGLCCTFVHRARSRYEHIPGPPRPSFLLGHLPCFWKKDEVGGRVLQDVFLDWAKKYGPVVRVNVFHKTSVIVTSPESVKKFLMSTKYNKDSKMYRALQTVFGERLFGQGLVSECNYERWHKQRRVIDLAFSRSSLVSLMETFNEKAEQLVEILEAKADGQTPVSMQDMLTYTAMDILAKAAFGMETSMLLGAQKPLSQAVKLMLEGITASRNTLAKFLPGKRKQLREVRESIRFLRQVGRDWVQRRREALKRGEEVPADILTQILKAEEGAQDDEGLLDNFVTFFIAGHETSANHLAFTVMELSRQPEIVARLQAEVDEVIGSKRYLDFEDLGRLQYLSQVLKESLRLYPPAWGTFRLLEEETLIDGVRVPGNTPLLFSTYVMGRMDTYFEDPLTFNPDRFGPGAPKPRFTYFPFSLGHRSCIGQQFAQMEVKVVMAKLLQRLEFRLVPGQRFGLQEQATLKPLDPVLCTLRPRGWQPAPPPPPC (s)-bicalutamide (S)-bicalutamide (+)-bicalutamide (S)-Casodex (2S)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(4-fluorophenyl)sulfonyl]-2-hydroxy-2-methylpropanamide small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) (r)-bicalutamide (R)-bicalutamide (-)-bicalutamide (R)-(-)bicalutamide (R)-Casodex (2R)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(4-fluorophenyl)sulfonyl]-2-hydroxy-2-methylpropanamide small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) (24s)-24-hydroxycholesterol (24S)-24-hydroxycholesterol (24S)-24-Hydroxycholesterol (24S)-hydroxycholesterol 24-Hydroxycholesterol 24S-hydroxy-cholesterol Cerebrosterol cholest-5-en-3beta,24S-diol Cholest-5-ene-3,24-diol (24S)-cholest-5-ene-3beta,24-diol small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) heme b Heme b small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) bicalutamide-cyp46a1-2 3581838 N/A 3581840 unspecified role unspecified role neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3581841 unspecified role unspecified role neutral component neutral component purified purified Q9Y6A2:p.Arg226Met mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.His81Ala 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 Q9Y6A2:p.Lys82Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Lys82Met mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Arg110Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Arg226Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Thr306Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Trp368Phe mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Ala474Val mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 3581838 FIGURES 1A, 1C, 8, 9 Changes elicited by racBIC in CYP46A1-imidazole adduct were identical to those in substrate-free CYP46A1. (s)_bicalutamide-cyp46a1-1 3581838 N/A 3581842 unspecified role unspecified role neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3581841 unspecified role unspecified role neutral component neutral component purified purified Q9Y6A2:p.Lys82Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.His81Ala mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Lys82Met mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Arg110Leu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Arg226Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Arg226Met mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Thr306Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Trp368Phe mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Ala474Val mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 3581838 FIGURES 1D, 8, 9 (r)_bicalutamide-cyp46a1-1 3581838 N/A 3581843 unspecified role unspecified role neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3581841 unspecified role unspecified role neutral component neutral component purified purified Q9Y6A2:p.Arg226Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Arg226Met mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Thr306Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Trp368Phe mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Ala474Val mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Lys82Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.His81Ala mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Lys82Met mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9Y6A2:p.Arg110Leu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 3581838 FIGURES 1E, 8, 9 (24s)_24_hydroxycholesterol-cyp46a1-1 3581838 N/A 3581841 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) N/A 3581844 unspecified role unspecified role neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction false false false 3581838 FIGURE 1B bicalutamide-cyp46a1-1 3581839 N/A 3581841 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false C-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-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 3581842 unspecified role unspecified role neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 N/A 3581843 unspecified role unspecified role neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 N/A 3581845 cofactor cofactor coenzyme neutral component neutral component purified purified chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 physical association physical association false false false TABLE 1, FIG. 3 2.1Å Rwork: 0.208 Rfree: 0.253