Udp gal

Manufactured by Promega

Sourced in United States, France

UDP-Gal is a nucleotide sugar that serves as a substrate for various glycosyltransferase enzymes involved in the synthesis of glycoproteins and glycolipids. It is a key intermediate in the Leloir pathway, which is responsible for the biosynthesis of galactose-containing oligosaccharides and polysaccharides.

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Lab products found in correlation

Synergy lx multi mode microplate reader, by Agilent Technologies (1 mentions) Udp glca, by Corning (1 mentions) Udp glo glycosyltransferase assay, by Promega (1 mentions)

Close spelling variants of this product

Ultra-pure UDP-Galactose (UDP-Gal)

3 protocols using udp gal

Characterization of GalS1 Glycosyltransferase

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All activated nucleotide sugars were purchased from CarboSource, Promega or Sigma. Screening of sugar-nucleotide donor specificities in the absence of acceptor substrate was done with the UDP-Glo glycosyltransferase assay (Promega) kit^{34 (link)}. Reactions (20 μl) consisted of 100 μM individual UDP-sugars (UDP-Gal, UDP-Arap, UDP-Xyl, UDP-Glc, UDP-GalA, UDP-GlcA, UDP-GlcNAc and UDP-GalNAc) and 4 μg of purified GalS1 in 50 mM HEPES and 100 mM NaCl (pH 7) at 30 °C for 18 h. The reaction mixture (5 μl) was mixed with an equal amount of UDP-Glo reagent in a 384-well assay plate (Corning 4513) and incubated for 1 h at room temperature before measuring luminescence using a Synergy LX Multi-mode microplate reader (BioTek). A standard curve was used for quantification of the UDP produced.
The quantity of UDP formed as a by-product of the GalT reaction was determined using the UDP-Glo glycosyltransferase assay (Promega) according to the manufacturer’s instructions, using either UDP-Gal (Promega) or UDP-Arap (CarboSource) as donor substrates. Standard GalT assays (20 μl) consisted of either UDP-Gal (250 μM) or UDP-Arap (400 μM) as activated nucleotide sugar donors, galactotetraose (400 μM) as an acceptor and 5 mM manganese(II) chloride in 50 mM HEPES (pH 7.0). Reactions were allowed to proceed at 30 °C for 2 h and the amount of UDP produced was determined as described above.

Prabhakar P.K., Pereira J.H., Taujale R., Shao W., Bharadwaj V.S., Chapla D., Yang J.Y., Bomble Y.J., Moremen K.W., Kannan N., Hammel M., Adams P.D., Scheller H.V, & Urbanowicz B.R. (2023). Structural and biochemical insight into a modular β-1,4-galactan synthase in plants. Nature plants, 9(3), 486-500.

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Nucleotide Sugar Hydrolysis Assay

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The ultra-pure UDP-Glc, UDP-Gal, and UDP-GlcA as well as UMP/CMP-Glo™ Glycosyltransferase Assay Kit were purchased from Promega Corporation, Madison, WI, USA. The measurement of the donor specificity of the recombinant PssA, determined as the hydrolysis of the nucleotide sugar derivative, was carried out by incubating a given amount (0.1–5 μg) of recombinant His6-PssA in the reaction buffer: 50 mM HEPES, 100 mM NaCl, 5 mM MgCl₂, 0.1% Tween-20 [23 (link)] with a given sugar substrate (100 μM) for 4 h at 28 °C. Control mixtures contained no enzyme. The hydrolysis reaction was stopped by adding UMP/CMP Detection Reagent^TM in a 1:1 ratio (25 μL:25 μL). After mixing, the plate was incubated at room temperature for 1 h. Then the level of luminescence was measured with Synergy H1 multi-detection reader (BioTek, Agilent Technologies, Santa Clara, CA, USA). The intensity of emitted light (luciferase reaction) is proportional to the amount of free UMP produced in the UDP-sugar hydrolysis reaction. Each reaction was performed in a separate well of a 96-well plate in two technical repeats.

Marczak M., Żebracki K., Koper P., Horbowicz A., Wójcik M, & Mazur A. (2023). A New Face of the Old Gene: Deletion of the PssA, Encoding Monotopic Inner Membrane Phosphoglycosyl Transferase in Rhizobium leguminosarum, Leads to Diverse Phenotypes That Could Be Attributable to Downstream Effects of the Lack of Exopolysaccharide. International Journal of Molecular Sciences, 24(2), 1035.

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Lipid Synthesis Protocol from Diverse Sources

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DAG (1-2-dioleoyl-sn-glycerol) was purchased from Avanti Polar Lipids (Alabama, US) and PG (L-α-Phosphatidyl-DL-glycerol ammonium salt from egg yolk lecithin) from Sigma-Aldrich (Saint-Quentin Fallavier, France). UDP-Gal was from Promega (Charbonnières-les-Bains, France).

Nitenberg M., Makshakova O., Rocha J., Perez S., Maréchal E., Block M.A., Girard-Egrot A, & Breton C. (2020). Mechanism of activation of plant monogalactosyldiacylglycerol synthase 1 (MGD1) by phosphatidylglycerol. Glycobiology, 30(6).

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