ChemicalBook CAS DataBase List UDP

UDP synthesis

8synthesis methods

Product Name:UDP
CAS Number:58-98-0
Molecular formula:C9H14N2O12P2
Molecular Weight:404.16

63-39-8 Synthesis

63-39-8
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58-98-0
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Yield:-

Reaction Conditions:

with tris(3-hydroxypropyl)phosphine;nicotinamide phosphoribosyltransferase;water;tris hydrochloride;magnesium chloride; pH=7.5 at 37;Kinetics;Enzymatic reaction;Reagent/catalyst;

Steps:

1 NAMPT Enzymatic Reactions
For the NTPase assays, NAMPT or NAMPT-H247A was incubated at 37 °C with an NTP in TMT buffer (50 mM Tris-HCl, 10 mM MgC12, 1 mM THP, pH 7.5). The routinely-used NTP concentration was 2 mM. However, for the kinetic analysis ATP and GTP hydrolysis, the NTP concentration ranged from 0.25 mM to 4 mM. Other agents (FK-866, CHS-828, GNI-50, PNP, NMN) were included where indicated. At the indicated times, an aliquot of the sample was quenched by the addition of an equal volume of 1 M perchloric acid (PCA). Samples were stored at -80 °C until processed for LC/MS/MS. The values for Vmax and Km (ATP and GTP hydrolysis) were deduced using the on-line Michaelis-Menten kinetics tool at (0160) http://www.graphpad.com/quickcalcs/ttestl/?Format=SEM For the NMN production assay, NAMPT (20 nM) was incubated in the presence of NAM (10 μΜ), PRPP (50 μΜ) in TMT buffer. Where indicated, NTPs (2 mM) were also included. NMN was assayed using a chemical method which converts NMN into a fluorescent derivative. Briefly, an aliquot (37.5 μ) of the NMN- containing sample was sequentially mixed with 15 μ of 20% acetophenone (in DMSO) and 15 μ of 2M KOH. The mixture was placed on ice for approximately 10 min. Next, 67.5 μ of 100% formic acid was added to each sample, vortexed, and then incubated at 37 °C for 20 min. Samples (100 μ) were transferred to a 96-well opaque bottom plate and fluorescence (Ex/Em = 382/445 nm) was measured using a SpectraMax M5 plate reader (Molecular Devices, Sunnyvale, CA).

References:

SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE;GARDELL, Stephen, J.;DISPAGNA, Mauro, Antonio WO2018/85379, 2018, A2 Location in patent:Paragraph 00128; 00133; 00134; 00136; 00138; 00140-00142