Active Recombinant Human Fucosyltransferase 8 (alpha (1, 6) fucosyltransferase), His-tagged

• Cat.No.: FUT8-577H
• Product Overview: Recombinanthuman FUT8 protein was expressed in Chinese Hamster Ovary cell line, CHO-derived;with an N-terminus human CD33 signal sequence, and 6-His tag. Asp32-Lys575.

Specification

• Species: Human
• Source: CHO
• Tag: His
• Protein Length: 32-575 a.a.
• Description: FUT8 is frequentlyfound at terminal sites on various glycols and is essential for thegeneration of many sugar epitomes. Well known FUT8 containing glycols includeLewis and ABO blood group antigens. Lewis epitomes are key elements involvedin leukocyte homing and extravasations process, thus are essential forlymphocyte maturation and natural defense functions.
• Form: Supplied as a 0.2 μmfiltered solution in Tris and NaCl.
• N-terminal Sequence: His
• Molecular Weight: 64 kDa
• Activity: Measured by itsability to hydrolyze the donor substrate GDP-fucose. The specific activity is> 0.75 pmole/min/μg, as measured under the described conditions.
• Endotoxin Level: <1.0 EU per 1 μgof the protein by the LAL method.
• SDS-PAGE: 58-62 kDa, reducingconditions
• Purity: >90%, by SDS-PAGEunder reducing conditions and visualized by Colloidal Coomassie Blue stain at5 μg per lane.
• Storage: Use amanual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -20 to -70 °C as supplied. 3months, -20 to -70 °C under sterile conditions after reconstitution.

Gene Information

• Gene Name: FUT8 fucosyltransferase 8 (alpha(1, 6) fucosyltransferase) [ Homo sapiens ]
• Official Symbol: FUT8
• Synonyms: FUT8; fucosyltransferase8 (alpha (1, 6) fucosyltransferase); alpha-(1, 6)-fucosyltransferase;OTTHUMP00000245300; OTTHUMP00000245301; OTTHUMP00000245302; OTTHUMP00000245303;OTTHUMP00000245304; OTTHUMP00000245305; OTTHUMP00000245306; OTTHUMP00000245307;OTTHUMP00000183477; OTTHUMP00000183478; OTTHUMP00000245299; EC 2.4.1.68; Fucosyltransferase8; GDP-L-Fuc: N-acetyl-beta-D-glucosaminide alpha 1, 6-fucosyltransferase;GDP-fucose-glycoprotein fucosyltransferase; Glycoprotein; 6-alpha-L-fucosyltransferase;alpha1-6FucT; MGC26465
• Gene ID: 2530
• mRNA Refseq: NM_004480
• Protein Refseq: NP_004471
• MIM: 602589
• UniProt ID: Q9BYC5
• Chromosome Location: 14q24.3
• Pathway: AsparagineN-linked glycosylation; Glycosaminoglycan biosynthesis-keratan sulfate;Metabolic pathways; Metabolism of proteins; N-Glycanbiosynthesis; N-glycan antennae elongation in the medial/trans-Golgi;N-glycan biosynthesis, complex type; N-glycan biosynthesis, complex type;Reactions specific to the complex N-glycan synthesis pathway; Transcriptionalmisregulation in cancer; Transport to the Golgi and subsequent modification
• Function: SH3domain binding; glycoprotein 6-alpha-L-fucosyltransferase activity;transferase activity, transferring glycosyl groups

Citation

A new approach for identifying positional isomers of glycans cleaved from monoclonal antibodies

Journal: The Analyst    PubMed ID: 34231555    Data: 2021/6/30

Authors: Irina Dyukova, Ahmed Ben Faleh, Thomas R. Rizzo

Article Snippet:The synthesis of G1(α1-6)F ( ) was performed in a total volume of 50 μL containing 0.27 mM of an acceptor G-NGA2 N-linked glycan (Dextra Laboratories, UK), 0.54 mM of GDP-Fuc (guanosine 5′-diphospho-β- l -fucose sodium salt) (Sigma-Aldrich), a MES (2-( N -morpholino)ethanesulfonic acid) buffer solution (100 mM, pH 7.0), and 0.12 mg mL ?1 of recombinant human α1,6-fucosyltransferase (FUT8) (Creative BioMart, USA).. The reaction was incubated overnight at 37 °C and then quenched by adding 30 μL of ice-cold acetonitrile to the mixture.The reaction was incubated overnight at 37 °C and then quenched by adding 30 μL of ice-cold acetonitrile to the mixture.

Positional isomers of G-NGA2 and G1F glycans with a scheme for chemoenzymatic synthesis of the G1(α1-6)F isomer using human α1,6-fucosyltransferase FUT8.

Arrival time distribution of G1F after 60 m SLIM-IMS separation ( i.e. , six separation cycles), representing a mixture of the G1(α1-6)F and G1(α1-3)F positional isomers.

ATDs of G1F glycan representing a mixture of G1(α1-6)F and G1(α1-3)F positional isomers (top) and the synthesised G1(α1-6)F isomer (bottom) after 60 m SLIM-IMS separation.