Adenosine-3′-phosphate-5′-phosphosulfate (PAPS), adenosine-3′,5′-diphosphate sodium salt (PAP), potassium phosphate monobasic, phenylmethylsulfonyl fluoride (PMSF), and dehydroepiandrosterone (DHEA) were purchased from Sigma-Aldrich (St. Louis, MO). PAPS was further purified (>98% as analyzed by HPLC) using a previously described procedure (Sekura 1981 ). Bacto tryptone and yeast extract were obtained from Becton Dickinson, Co. (Sparks, MD) and Hydroxyapatite (Bio-Gel HT) from Bio-Rad Laboratories (Hercules, CA). Tween 20 was purchased from J.T. Baker Chemicals (Phillipsburg, NJ). Isopropyl-1-thio-D-galactopyranoside (IPTG) was from AMRESCO Inc. (Solon, OH). All other chemicals used were of the highest purity commercially available.
Adenosine 3 phosphate 5 phosphosulfate
Manufactured by Merck Group
Sourced in Belgium, United States
Adenosine-3'-phosphate-5'-phosphosulfate is a chemical compound that functions as a cofactor in various enzymatic reactions. It is a key intermediate in the biosynthesis of sulfated biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
Dehydroepiandrosterone, by Merck Group (1 mentions)
Bacto tryptone and yeast extract, by BD (1 mentions)
Hydroxyapatite bio gel ht, by Bio-Rad (1 mentions)
Phenylmethylsulfonyl fluoride, by Merck Group (1 mentions)
Potassium phosphate monobasic, by Merck Group (1 mentions)
Udp α d glucuronic acid, by Merck Group (1 mentions)
Udp α d galactose, by Merck Group (1 mentions)
Udp α d glucose, by Merck Group (1 mentions)
Udp β l arabinofuranose, by Peptide Institute (1 mentions)
Cmp n acetylneuraminic acid, by Merck Group (1 mentions)
Verapamil, by Merck Group (1 mentions)
Diclofenac sodium salt, by Merck Group (1 mentions)
Indinavir, by Merck Group (1 mentions)
Sildenafil, by Merck Group (1 mentions)
Enalapril, by Merck Group (1 mentions)
Terfenadine, by Merck Group (1 mentions)
Rifabutin, by Merck Group (1 mentions)
Ramipril, by Merck Group (1 mentions)
Atorvastatin, by Merck Group (1 mentions)
Quinidine, by Merck Group (1 mentions)
5 protocols using adenosine 3 phosphate 5 phosphosulfate
1
PAPS Purification and Reagent Procurement
2
Enzymatic Synthesis of Glycosidic Substrates
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Substrates were obtained from the following sources: UDP-α-d-xylose, UDP-β-l-arabinopyranose and UDP-α-d-galacturonic acid (Carbosource Services, Complex Carbohydrate Research Center, Athens, GA); UDP-α-d-glucuronic acid, UDP-α-d-glucose, UDP-α-d-galactose, UDP-N-acetyl-α-d-glucosamine, UDP-N-acetyl-α-d-galactosamine, GDP-α-d-mannose, GDP-β-l-fucose, GDP-α-d-glucose, adenosine 3′-phosphate 5′ phosphosulfate, CMP-N-acetylneuraminic acid and ADP-α-d-glucose (Sigma-Aldrich, St Louis, MO); and UDP-β-l-arabinofuranose (Peptides International, Louisville, KY). UDP-β-l-rhamnose was enzymatically synthesised as described earlier (18 (link)).
3
Characterization of Drug Metabolism Assays
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Uridine 59-diphosphoglucuronic acid trisodium salt (UDPGA), NADPH, alamethicin from Trichoderma viride, adenosine 3-phosphate 5-phosphosulfate, diclofenac sodium salt, midazolam hydrochloride, felodipine, S-(+)-mephenytoin, raloxifene hydrochloride, lovastatin, nisoldipine, nifedipine, methadone, quinidine, testosterone, benzydamine hydrochloride, cisapride, cyclosporin, terbutaline, sildenafil, verapamil, trazodone, atorvastatin, simvastatin, buspirone, rifabutin, saquinavir, terfenadine, zolpidem, repaglinide, indinavir, alprazolam, carbazeran, enalapril, ramipril , and dabigatran etexilate were purchased from Sigma-Aldrich (St. Louis, MO). NADPH-regenerating system containing NADP + , glucose-6phosphate, and glucose-6-phosphate dehydrogenase was purchased from Corning (Wiesbaden, Germany). Organic solvents were of LC-mass spectrometry (MS) or higher quality grade and were acquired from VWR International (Radnor, PA) or Fisher Scientific UK Ltd (Loughborough, UK). Purified water was obtained from Milli-Q Integral 5 Water Purification System (Merck KGaA, Darmstadt, Germany). Phosphate buffer, HQM Hepatocyte/Enterocyte Incubation Medium, and Co-Factor N for MetMax were purchased from In Vitro ADMET Laboratories (Columbia, MD).
4
Microsomal and Cytosolic Metabolism Study
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Pooled human liver microsomes (HLM; n=50, mixed gender) and pooled human liver cytosol (HLCYT, mix gender) were purchased from Tebu Bio (Boechout, Belgium) and Sigma-Aldrich (Bornem, Belgium), respectively. RDP standard (98% purity) was obtained from TRC (Toronto, Canada); reduced β-nicotinamide adenine dinucleotide 2'-phosphate (NADPH; purity>95%), uridine 5'-diphosphoglucuronic acid (UDPGA; purity>98%), adenosine 3'phosphate 5'-phosphosulfate (PAPS; purity>96%), DPHP and TPHP-d 15 (used as internal standard, IS) were purchased from Sigma-Aldrich (Bornem, Belgium). The standard TPHP was purchased from Chiron AS (Trondheim, Norway). The DPHP-d 10 standard (also used as IS) was custom synthesized by Dr. Vladimir Belov (Max Planck Institute, Göttingen, Germany).
Acetonitrile, methanol and ammonium acetate were purchased from Merck KgA Chemicals (Darmstadt, Germany). para-HO-TPHP was synthesized by the Max Planck Institute (Göttingen, Germany) with a purity of 98%. meta-HO-TPHP was synthesized by the Organic Chemistry Synthesis Group from Duke University (NC, USA) and kindly donated by prof. Heather Stapleton (Duke University, NC, USA). Ultrapure water (18.2 MΩ) was obtained using an Elga LabWater water purification instrument (Saint Maurice, France). Oasis WAX cartridges (3 cc, 60 mg, 30 µm) were obtained from Waters (Milford, Massachusetts, USA).
Acetonitrile, methanol and ammonium acetate were purchased from Merck KgA Chemicals (Darmstadt, Germany). para-HO-TPHP was synthesized by the Max Planck Institute (Göttingen, Germany) with a purity of 98%. meta-HO-TPHP was synthesized by the Organic Chemistry Synthesis Group from Duke University (NC, USA) and kindly donated by prof. Heather Stapleton (Duke University, NC, USA). Ultrapure water (18.2 MΩ) was obtained using an Elga LabWater water purification instrument (Saint Maurice, France). Oasis WAX cartridges (3 cc, 60 mg, 30 µm) were obtained from Waters (Milford, Massachusetts, USA).
5
Quantifying Hepatic Metabolism of 5Cl-THJ-018
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5Cl-THJ-018 was acquired from Cayman Chemical (Ann Arbor, MI, USA). HLMs (mixed gender, n=50) were acquired from Tebu-Bio (Boechout, BE). HLCYT (mixed gender, n=150), theophylline (anhydrous, > 99 %), 2,5-uridinediphosphate glucuronic acid (UDPGA), adenosine-3'-phosphate 5'phosphosulfate (PAPS, > 60 %) lithium salt hydrate, alamethicin (neat, > 98 %), dimethyl sulfoxide (DMSO), and 4-nitrophenol (4-NP) were obtained from Sigma-Aldrich (Missouri, US). NADPH tetrasodium salt hydrate (> 96 %) was purchased from Acros (Geel, BE). Acetonitrile (ACN, HPLC-grade) and methanol (MeOH, ≥ 99.9 % LC-MS grade) were acquired from Fisher Chemical (Loughborough, GB), formic acid (> 98 %) and hydrochloric acid (37%) from Merck KGaA (Darmstadt, DE). A 100 mM TRIS-buffer was prepared by dissolving 12.11 g Trizma base (Janssen Chimica, Beerse, BE) and 1.02 g MgCl2 (Merck KGaA, Darmstadt, DE) in 1 L ultrapure water. The pH was adjusted to 7.4 by adding 1 M HCl solution. Ultra-pure water was produced in-house with a PURELAB-purifier system of Elga Labwater (Tienen, BE).
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