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Gdp fucose

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GDP-fucose is a nucleotide sugar that serves as a substrate for various glycosyltransferase enzymes involved in the synthesis of fucosylated glycans. It is a key component in the biosynthesis of complex carbohydrates and plays a role in various biological processes.

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Lab products found in correlation

Human serum albumin (hsa), by Merck Group (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Ftvii, by R&D Systems (1 mentions)

5 protocols using gdp fucose

1

FUT8 Enzyme Fucosylation Experiments

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All the FUT8 in vitro experiments were carried out for the indicated times in 0.1 M of MES buffer (pH 7) at 37 °C. For the reaction with glycoproteins, 1 μmol of the substrates (EPO-HM, EPO-PM, and EPO-GlcNAc) were mixed with 1 mM of GDP-Fucose (Biosynth Carbosynth) and 0.05 mg/mL of FUT8. For chemically modified N-glycans and glycopeptides, the reactions were performed with 5 molar equivalents of GDP-Fucose and 0.5 mg/mL of the FUT8 enzyme.
Zhang R., Yang Q., Boruah B.M., Zong G., Li C., Chapla D., Yang J.Y., Moremen K.W, & Wang L.X. (2021). Appropriate aglycone modification significantly expands the glycan substrate acceptability of α1,6-fucosyltransferase (FUT8). The Biochemical journal, 478(8), 1571-1583.
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2

Fucosyltransferase VII Treatment of Tregs

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Cells were treated with fucosyltransferase VII (FTVII) or buffer-only at 37 °C for 60 minutes. Tregs were resuspended at 3 × 106 cells/30 ul in HBSS with 2 ug FTVII (RnD Systems), 10 mM Hepes, 0.1% Human Serum Albumin, and 1 mM GDP fucose (Carbosynth). Treatment success was determined by flow cytometry with HECA452 mAb (Biolegend).
Donnelly C., Dykstra B., Mondal N., Huang J., Kaskow B.J., Griffin R., Sackstein R, & Baecher-Allan C. (2018). Optimizing human Treg immunotherapy by Treg subset selection and E-selectin ligand expression. Scientific Reports, 8, 420.
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3

MSC Fucosylation Protocol

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MSCs were harvested, washed 2x with PBS, and resuspended at 2×107 cells/ml in FTVI reaction buffer, containing 20mM HEPES (Gibco), 0.1% human serum albumin (Sigma), 1mM GDP-fucose (Carbosynth), and 60 μg/ml purified FTVI enzyme in Hank’s Balanced Salt Solution (HBSS). Cells were incubated at 37°C for 1 hour. For some experiments, “buffer only” controls were performed in an identical fashion but excluding the FTVI enzyme and GDP-fucose from the reaction. After the reaction, the cells were washed 2x with PBS and used immediately for downstream experiments.
Dykstra B., Lee J., Mortensen L.J., Yu H., Wu Z.L., Lin C.P., Rossi D.J, & Sackstein R. (2016). Glycoengineering of E-selectin ligands by intracellular versus extracellular fucosylation differentially affects osteotropism of human mesenchymal stem cells. Stem cells (Dayton, Ohio), 34(10), 2501-2511.
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4

Fucosylation of CD19-CAR T cells

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CD19‐CAR T cells expanded for 9 days either with IL‐2‐ or IL‐7/IL‐15 were treated on Hanks' Balanced Salt solution (0.1% human serum albumin and 10 mM HEPES) with GDP‐fucose (Biosynth Carbosynth, Compton, UK) and FTVII (RD Systems). One million cells were incubated in 20 μL of buffer containing 1 mM of GDP‐fucose and 70 μg/mL of purified FTVII enzyme at 37°C for 1 h as previously detailed.17 Control cells were incubated in the same solution but without FTVII/GDP‐fucose (buffer‐treated [BT] cells). After the enzymatic reaction, cells were always washed twice with PBS before downstream experiments.
Sánchez‐Martínez D., Gutiérrez‐Agüera F., Romecin P., Vinyoles M., Palomo M., Tirado N., Zanetti S.R., Juan M., Carlet M., Jeremias I, & Menéndez P. (2021). Enforced sialyl‐Lewis‐X (sLeX) display in E‐selectin ligands by exofucosylation is dispensable for CD19‐CAR T‐cell activity and bone marrow homing. Clinical and Translational Medicine, 11(2), e280.
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5

Fucosylation Enhancement in Cell Cultures

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Cells were treated with 60 mU/mL fucosyltransferase-VI (FTVI (Warrior Therapeutics, Sudbury, MA), in HBSS containing 20 mM HEPES, 0.1% human serum albumin (HSA, Sigma) and 1 mM GDP-fucose (Carbosynth, Compton, UK) for 45 min at 37°C. Control consisted of cells treated with buffer containing all components except FTVI enzyme. The efficacy of the FTVI treatment was confirmed by evaluating for increased expression of E-selectin ligands as assessed by both flow cytometry and western blot analysis using E-Ig chimera as probe.
Videira P.A., Silva M., Martin K.C, & Sackstein R. (2018). Ligation of the CD44 Glycoform HCELL on Culture-Expanded Human Monocyte-derived Dendritic Cells Programs Transendothelial Migration. Journal of immunology (Baltimore, Md. : 1950), 201(3), 1030-1043.
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