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dc.contributor.authorAl-Amoodi, Asma S.
dc.contributor.authorSakashita, Kosuke
dc.contributor.authorAli, Amal J.
dc.contributor.authorZhou, Ruoyu
dc.contributor.authorLee, Jae Man
dc.contributor.authorTehseen, Muhammad
dc.contributor.authorLi, Mo
dc.contributor.authorIzpisua Belmonte, Juan Carlos
dc.contributor.authorKusakabe, Takahiro
dc.contributor.authorMerzaban, Jasmeen
dc.date.accessioned2020-09-17T09:08:48Z
dc.date.available2020-09-17T09:08:48Z
dc.date.issued2020-09-09
dc.identifier.citationAl-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.0c00523
dc.identifier.issn0006-2960
dc.identifier.issn1520-4995
dc.identifier.doi10.1021/acs.biochem.0c00523
dc.identifier.urihttp://hdl.handle.net/10754/665213
dc.description.abstractRecruitment 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.
dc.publisherAmerican Chemical Society (ACS)
dc.relation.urlhttps://pubs.acs.org/doi/10.1021/acs.biochem.0c00523
dc.rightsThis document is the Accepted Manuscript version of a Published Work that appeared in final form in Biochemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/10.1021/acs.biochem.0c00523.
dc.titleUsing Eukaryotic Expression Systems to Generate Human α1,3 Fucosyltransferases that Effectively Create Selectin-binding Glycans on Stem Cells
dc.typeArticle
dc.contributor.departmentBioscience Program
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentProteomics and Protein Expression
dc.identifier.journalBiochemistry
dc.rights.embargodate2021-09-09
dc.eprint.versionPost-print
dc.contributor.institutionCurrent Address: Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, 9420 Athena Circle Drive, La Jolla, CA, 92037 USA.
dc.contributor.institutionLaboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Motooka 744, Nishi-ku, Fukuoka 819-0395, Japan.
dc.contributor.institutionGene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
kaust.personAl-Amoodi, Asma
kaust.personSakashita, Kosuke
kaust.personAli, Amal J.
kaust.personTehseen, Muhammad
kaust.personLi, Mo
kaust.personMerzaban, Jasmeen S.
refterms.dateFOA2020-09-17T09:09:26Z
dc.date.published-online2020-09-09
dc.date.published-print2020-10-06


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