The binding of nucleosides to LF-rBmpD was monitored with MST (43 (link)). Adenosine, guanosine, inosine, xanthosine (Sigma-Aldrich, Darmstadt, Germany), and ribose (negative-control ligand; Sigma-Aldrich) were mixed with LF-rBmpD (final concentration, 500 nM) in a 24-point serial dilution. The concentration of the ligands ranged from 5 mM to 1.2 nM. Samples were filled into zero-background standard-treated capillaries (product number MO-AZ002; NanoTemper Technologies, Munich, Germany) and were measured with Monolith.NT115 LabelFree equipment (NanoTemper Technologies), using 60% light-emitting diode (LED) power and medium MST power. The data were analyzed by MO.Affinity Analysis software (NanoTemper Technologies) and GraphPad Prism (version 8.0; GraphPad Software, San Diego, CA, USA). No dissociation constants are displayed because results of only one experiment are shown.
Xanthosine
Manufactured by Merck Group
Sourced in United States
Xanthosine is a laboratory reagent primarily used in biochemical and molecular biology research. It is a nucleoside composed of the purine base xanthine and the sugar ribose. Xanthosine serves as a precursor for the synthesis of other nucleotides and can be used as a substrate in various enzymatic reactions.
Automatically generated - may contain errors
Lab products found in correlation
Inosine, by Merck Group (2 mentions)
Creatinine, by Merck Group (2 mentions)
Pseudouridine, by Merck Group (2 mentions)
N6 methyladenosine, by Merck Group (2 mentions)
1 methyladenosine, by Merck Group (2 mentions)
Cytidine, by Merck Group (2 mentions)
Ribose, by Merck Group (1 mentions)
Guanosine, by Merck Group (1 mentions)
Adenosine, by Merck Group (1 mentions)
Mo affinity analysis software, by NanoTemper (1 mentions)
Cortisone, by Merck Group (1 mentions)
Cortisol, by Merck Group (1 mentions)
Xanthurenic acid, by Merck Group (1 mentions)
Lc ms grade, by Thermo Fisher Scientific (1 mentions)
Xanthine, by Merck Group (1 mentions)
Kynurenic acid, by Merck Group (1 mentions)
L acetylcarnitine, by Merck Group (1 mentions)
Taurine, by Merck Group (1 mentions)
Hypoxanthine, by Merck Group (1 mentions)
Debrisoquine sulfate, by Merck Group (1 mentions)
5 protocols using xanthosine
1
Nucleoside Binding Affinity to LF-rBmpD
2
LC-MS Quantification of Metabolites
3
Capillary-Based Silica Functionalization for Nucleoside Analysis
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Fused-silica capillary (250 μm i.d. × 360 μm o.d.) was purchased from Yongnian Optic Fiber Plant (Hebei, China). Tetramethoxysilane (TMOS) and 3-mercaptopropyltrimethoxysilane (MPTMS) were purchased from Wuhan University Silicone New Material (Wuhan, China). Azobisisobutyronitrile (AIBN) and poly(ethylene glycol) with the molecular weight of 6000 (PEG-6000) were all purchased from Shanghai Chemical Reagent Corporation (Shanghai, China). AIBN was purified by recrystallization from ethanol at 40°C. 3-acrylamidophenylboronic acid (AAPBA) and creatinine were purchased from Sigma-Aldrich (Beijing, China). Organic solvents were all of HPLC grade. The water used throughout all experiments was purified using a Milli-Q apparatus (Millipore, Bradford, USA). All other reagents were obtained from various commercial sources and were of analytical grade unless otherwise indicated.
2′-Deoxycytidine (dC), 2′-deoxyguanosine (dG), 2′-deoxyadenosine (dA), thymidine (T), cytidine (rC), guanosine (rG), adenosine (rA), uridine (rU), 1-methyladenosine, N6-methyladenosine, 5′-deoxyadenosine, inosine, xanthosine, 3-methylcytidine, N4-acetylcytidine, 5-methyluridine, 3-methyluridine, pseudouridine, double hydrogen zeatin-riboside (DHzR) were purchased from Sigma-Aldrich (Beijing, China). The standard solution of each analyte was prepared at 1.0 mg/mL in H2O and stored at −20°C.
2′-Deoxycytidine (dC), 2′-deoxyguanosine (dG), 2′-deoxyadenosine (dA), thymidine (T), cytidine (rC), guanosine (rG), adenosine (rA), uridine (rU), 1-methyladenosine, N6-methyladenosine, 5′-deoxyadenosine, inosine, xanthosine, 3-methylcytidine, N4-acetylcytidine, 5-methyluridine, 3-methyluridine, pseudouridine, double hydrogen zeatin-riboside (DHzR) were purchased from Sigma-Aldrich (Beijing, China). The standard solution of each analyte was prepared at 1.0 mg/mL in H2O and stored at −20°C.
4
LC-MS Metabolite Quantification Protocol
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The solvents methanol and acetonitrile at LC-MS grade were purchased from Scharlab (Barcelona, Spain), as well as the eluent additive formic acid for LC-MS and reagent grade ammonium acetate. HPLCgrade water was obtained with Milli-Q water purification system (Millipore Ltd., Bedford, MA, USA). Leucine-enkephalin HPLC-grade (massaxis recalibration) and analytical standards of riboflavin, cytidine, 1methylnicotinamide, xanthosine, asymmetric dimethylarginine and docosahexaenoic acid were purchased werewas purchased from Sigma-Aldrich (Saint Louis, MO, USA).
5
Quantification of Urinary Methylated Nucleosides
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Starting point. Starting from the results of our already published studies from 2006, 2007 and 2014 (9-11) , the spectrum of mNS measured in urine was supplemented by hypoxanthosine, L-tryptophan and guanosine (Table I). In total, 15 mNS and the sum of all measured mNS were quantified in the urine of study participants.
Reagents. The following mNS standards were purchased from Sigma (St Louis, MO, USA): pseudouridine, uridine, cytidine, 1-methyladenosine, 5-methylcytidine, 1-methylinosine, guanosine, xanthosine, 1-methylguanosine, 2-methylguanosine, N2,N2dimethylguanosine, N6-methyladenosine, hypoxanthosine and Ltryptophan. All of them were of HPLC purity. Ammonium dihydrogen phosphate buffer (NH 4 H 2 PO 4 ), methanol and acetonitrile were obtained from Baker (Phillipsburg, NJ, USA) of Baker-analyzed HPLC grade. Deionized water was acquired from a Milli O plus purification system (Millipore, Bedford, MA, USA).
Reagents. The following mNS standards were purchased from Sigma (St Louis, MO, USA): pseudouridine, uridine, cytidine, 1-methyladenosine, 5-methylcytidine, 1-methylinosine, guanosine, xanthosine, 1-methylguanosine, 2-methylguanosine, N2,N2dimethylguanosine, N6-methyladenosine, hypoxanthosine and Ltryptophan. All of them were of HPLC purity. Ammonium dihydrogen phosphate buffer (NH 4 H 2 PO 4 ), methanol and acetonitrile were obtained from Baker (Phillipsburg, NJ, USA) of Baker-analyzed HPLC grade. Deionized water was acquired from a Milli O plus purification system (Millipore, Bedford, MA, USA).
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