ChEMBL ligand: CHEMBL511275
L-Tryptophan hydroxylase 1/Tryptophan 5-hydroxylase 1 in Human [ChEMBL: CHEMBL5689] [GtoPdb: 1241] [UniProtKB: P17752] L-Tryptophan hydroxylase 1/Tryptophan 5-hydroxylase 1 in Rat [ChEMBL: CHEMBL4809] [GtoPdb: 1241] [UniProtKB: P09810] There should be some charts here, you may need to enable JavaScript!
L-Tryptophan hydroxylase 2/Tryptophan 5-hydroxylase 2 in Human [ChEMBL: CHEMBL5433] [GtoPdb: 1242] [UniProtKB: Q8IWU9] There should be some charts here, you may need to enable JavaScript!

DB	Assay description	Assay Type	Standard value	Standard parameter	Original value	Original units	Original parameter	Reference
L-Tryptophan hydroxylase 1/Tryptophan 5-hydroxylase 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5689] [GtoPdb: 1241] [UniProtKB: P17752]
ChEMBL	In Vitro Enzyme Activity Assay: The full-length coding sequences of human TPH1 and TPH2 were PCR amplified, ligated into a MBP fusion vector (pMalc2x, New England Biolabs, MA, USA) and transformed into SCS1 (Stratagene, CA, USA) to amplify plasmid DNA. For the overexpression of TPH proteins, the constructs were transformed into Rosetta (DE3) (Novagen®/EMD Millipore, Mass., USA) and cultivated in terrific broth (TB) medium (AppliChem, Darmstadt, Germany) at 37° C. When the bacterial cultures reached an ≈2, expression was induced with 0.5 mM IPTG (AppliChem, Darmstadt, Germany) over night at 17° C. The purification of soluble proteins started with sonication-mediated cell disruption in lysis buffer (1×PBS pH 7.4, 0.5 M NaCl, 5% Glycerol+CHAPS, DTT, PMSF, benzonase), followed by affinity purification (MBPTrap, GE Healthcare, UK) and gel filtration (26/60 Superdex 200 prep grade, GE Healthcare, UK), according to the manufacturer's protocol. The quality of protein expression and solubility was controlled by SDS-PAGE and Coomassie blue staining.The enzymatic reaction was carried out in black 96-well flat bottom plates (Corning GmbH, Wiesbaden). TPH1 and TPH2 activities were measured in a reaction mixture containing 50 mM 4-Morpholineethanesulfonic acid (MES), pH 7.0, 40 μM tryptophan, 200 mM ammonium sulfate, 25 μM ferrous ammonium sulfate, 50 μM tetrahydrobiopterin, 25 μg/ml catalase, and 7 mM DTT. The reactions were initiated by adding TPH1 or TPH2 to a final concentration of 5 μg/ml. Initial velocity of the reactions was determined by following the change of fluorescence at 330 nm (excitation wavelength=300 nm) (Infinite M200, Tecan, Crailsheim).	B	5.7	pIC50	2010	nM	IC50	US-10683309-B2. Xanthine derivatives, their use as a medicament, and pharmaceutical preparations comprising the same (2020)
GtoPdb	-	-	6.23	pIC50	583	nM	IC50	J Med Chem (2021) 64: 1037-1053 [PMID:33417443]
ChEMBL	Inhibition of Tryptophan hydroxylase 1 (unknown origin) incubated for 4 hrs by fluorescence based assay	B	6.23	pIC50	583	nM	IC50	J Med Chem (2021) 64: 1037-1053 [PMID:33417443]
L-Tryptophan hydroxylase 1/Tryptophan 5-hydroxylase 1 in Rat (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL4809] [GtoPdb: 1241] [UniProtKB: P09810]
ChEMBL	Inhibition of TPH-1-mediated serotonin biosynthesis in rat RBL2H3 cells after 48 hrs by RP-HPLC analysis	B	6.4	pIC50	400	nM	IC50	J Med Chem (2014) 57: 4692-4709 [PMID:24844139]
L-Tryptophan hydroxylase 2/Tryptophan 5-hydroxylase 2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5433] [GtoPdb: 1242] [UniProtKB: Q8IWU9]
ChEMBL	In Vitro Enzyme Activity Assay: The full-length coding sequences of human TPH1 and TPH2 were PCR amplified, ligated into a MBP fusion vector (pMalc2x, New England Biolabs, MA, USA) and transformed into SCS1 (Stratagene, CA, USA) to amplify plasmid DNA. For the overexpression of TPH proteins, the constructs were transformed into Rosetta (DE3) (Novagen®/EMD Millipore, Mass., USA) and cultivated in terrific broth (TB) medium (AppliChem, Darmstadt, Germany) at 37° C. When the bacterial cultures reached an ≈2, expression was induced with 0.5 mM IPTG (AppliChem, Darmstadt, Germany) over night at 17° C. The purification of soluble proteins started with sonication-mediated cell disruption in lysis buffer (1×PBS pH 7.4, 0.5 M NaCl, 5% Glycerol+CHAPS, DTT, PMSF, benzonase), followed by affinity purification (MBPTrap, GE Healthcare, UK) and gel filtration (26/60 Superdex 200 prep grade, GE Healthcare, UK), according to the manufacturer's protocol. The quality of protein expression and solubility was controlled by SDS-PAGE and Coomassie blue staining.The enzymatic reaction was carried out in black 96-well flat bottom plates (Corning GmbH, Wiesbaden). TPH1 and TPH2 activities were measured in a reaction mixture containing 50 mM 4-Morpholineethanesulfonic acid (MES), pH 7.0, 40 μM tryptophan, 200 mM ammonium sulfate, 25 μM ferrous ammonium sulfate, 50 μM tetrahydrobiopterin, 25 μg/ml catalase, and 7 mM DTT. The reactions were initiated by adding TPH1 or TPH2 to a final concentration of 5 μg/ml. Initial velocity of the reactions was determined by following the change of fluorescence at 330 nm (excitation wavelength=300 nm) (Infinite M200, Tecan, Crailsheim).	B	5.65	pIC50	2230	nM	IC50	US-10683309-B2. Xanthine derivatives, their use as a medicament, and pharmaceutical preparations comprising the same (2020)

ChEMBL data shown on this page come from version 36:

Zdrazil B, Felix E, Hunter F, Manners EJ, Blackshaw J, Corbett S, de Veij M, Ioannidis H, Lopez DM, Mosquera JF, Magarinos MP, Bosc N, Arcila R, Kizilören T, Gaulton A, Bento AP, Adasme MF, Monecke P, Landrum GA, Leach AR. (2024). The ChEMBL Database in 2023: a drug discovery platform spanning multiple bioactivity data types and time periods. Nucleic Acids Res., 52(D1). DOI: 10.1093/nar/gkad1004. [EPMCID:10767899] [PMID:37933841]
Davies M, Nowotka M, Papadatos G, Dedman N, Gaulton A, Atkinson F, Bellis L, Overington JP. (2015) 'ChEMBL web services: streamlining access to drug discovery data and utilities.' Nucleic Acids Res., 43(W1). DOI: 10.1093/nar/gkv352. [EPMCID:25883136]