Using Eukaryotic Expression Systems to Generate Human α1,3 Fucosyltransferases that Effectively Create Selectin-binding Glycans on Stem Cells
Type
ArticleAuthors
Al-Amoodi, Asma S.
Sakashita, Kosuke

Ali, Amal J.

Zhou, Ruoyu
Lee, Jae Man
Tehseen, Muhammad

Li, Mo

Izpisua Belmonte, Juan Carlos
Kusakabe, Takahiro
Merzaban, Jasmeen

KAUST Department
Bioscience ProgramBiological and Environmental Sciences and Engineering (BESE) Division
Proteomics and Protein Expression
Date
2020-09-09Online Publication Date
2020-09-09Print Publication Date
2020-10-06Embargo End Date
2021-09-09Permanent link to this record
http://hdl.handle.net/10754/665213
Metadata
Show full item recordAbstract
Recruitment of circulating cells towards target sites is primarily dependent on selectin/ligand adhesive interactions.Glycosyltransferases are involved in the creation of selectin ligands on proteins and lipids. α1,3-fucosylation is imperative for the creation of selectin ligands and a number of fucosyltransferases (FTs) are able to modify terminal lactosamines on cells to create these ligands. One FT, fucosyltransferase VI (FTVI) adds a fucose in an α1,3 configuration to N-acetylglucosamine to generate sialyl Lewis X (sLex) epitopes on proteins of live cells and enhances their ability to bind E-selectin. Although a number of recombinant human FTVIs have been purified, apart from limited commercial enzymes, they were not characterized for their activity on live cells. Here we focused on establishing a robust method to produce FTVI that is active on living cells (hematopoietic cells and mesenchymal stromal cells). To this end, we used two expression systems, Bombyx mori (silkworm) and Pichia pastoris (yeast), to produce significant amounts of N-terminally tagged FTVI and demonstrated that these enzymes have superior activity when compared to currently available commercial enzymes that are produced from various expression systems. Overall, we outline a scheme to attain large amounts of highly active FTVI that can be used for the applications of FTVI in enhancing the engraftment of cells lacking the sLex epitopes.Citation
Al-Amoodi, A. S., Sakashita, K., Ali, A. J., Zhou, R., Lee, J. M., Tehseen, M., … Merzaban, J. S. (2020). Using Eukaryotic Expression Systems to Generate Human α1,3 Fucosyltransferases that Effectively Create Selectin-binding Glycans on Stem Cells. Biochemistry. doi:10.1021/acs.biochem.0c00523Publisher
American Chemical Society (ACS)Journal
BiochemistryAdditional Links
https://pubs.acs.org/doi/10.1021/acs.biochem.0c00523ae974a485f413a2113503eed53cd6c53
10.1021/acs.biochem.0c00523