Enzymatic characterization of bovine alpha-1,3-galactosyltransferase. Validation of a radiometric assay and kinetic mechanism
2005 (Spanish)In: Afinidad, ISSN 0001-9704, Vol. 62, no 519, 505-512 p.Article in journal (Refereed) Published
alpha 3-Galactosyltransferase (alpha 3GT) transfers galactose from UDP-Gal (sugar nucleotide donor) to the N-acetyllactosaminyl or lactosyl terminal groups of glycoproteins and glycolipids, catalyzing the formation of an alpha-1,3 glycosidic bond. The terminal saccharide Gal alpha 3NAcGal beta 4Glu-R is the main antigenic determinant responsible of the hyperaccute rejection in xenotransplantation. A radiometric assay for the determination of alpha 3GT activity is implemented and validated. The recombinant enzyme (catalytic domain) expressed in Eschericia coli follows a bi bi sequential ordered kinetic mechanism with binding of donor substrate (UDP-Gal) first and acceptor substrate to form a productive ternary complex. K-M values are 30 mu M for UDP-Gal, and 1.2 mM for lactose.
Place, publisher, year, edition, pages
2005. Vol. 62, no 519, 505-512 p.
galactosyltransferase, radiometric assay, activity, kinetic mechanism, sequential ordered mechanim, validation, histo-blood group, glycosyl transferases, alpha, specificity, udp-galactose-beta-galactoside-alpha-1,3-galactosyltransferase, galactosyltransferase, glycosyltransferases, recombinant, hydrolases, sequences
IdentifiersURN: urn:nbn:se:kth:diva-15335ISI: 000237566100025OAI: oai:DiVA.org:kth-15335DiVA: diva2:333376
QC 201005252010-08-052010-08-05Bibliographically approved