Exploration of the active site of β4GalT7: modifications of the aglycon of aromatic xylosides

• Published in: Organic & biomolecular chemistry Vol. 13; no. 11; pp. 3351 - 3362
• Main Authors: Siegbahn, Anna, Thorsheim, Karin, Stahle, Jonas, Manner, Sophie, Hamark, Christoffer, Persson, Andrea, Tykesson, Emil, Mani, Katrin, Westergren-Thorsson, Gunilla, Widmalm, Goran, Ellervik, Ulf
• Format: Journal Article
• Language: English
• Published: CAMBRIDGE Royal Soc Chemistry 01.01.2015
• Subjects: Alcohols - chemistry; Basic Medicine; Catalytic Domain; Cell and Molecular Biology; Cell- och molekylärbiologi; Chemistry; Chemistry, Organic; Galactosyltransferases - chemistry; Galactosyltransferases - metabolism; Glycosides - chemical synthesis; Glycosides - chemistry; Glycosides - metabolism; Humans; Läkemedelskemi; Medical and Health Sciences; Medicin och hälsovetenskap; Medicinal Chemistry; Medicinska och farmaceutiska grundvetenskaper; Molecular Docking Simulation; Physical Sciences; Science & Technology; Tumor Cells, Cultured; UDP-GALACTOSE; CHONDROITIN SULFATE BIOSYNTHESIS; GLYCOSAMINOGLYCAN BIOSYNTHESIS; INHIBITION; BETA-D-XYLOSIDES; LINKAGE REGION; DOCKING; HEPARAN-SULFATE; TUMOR ATTENUATION; PROTEOGLYCAN
• ISSN: 1477-0520; 1477-0539; 1477-0539
• DOI: 10.1039/c4ob02632b

Proteoglycans (PGs) are macromolecules that consist of long linear polysaccharides, glycosaminoglycan (GAG) chains, covalently attached to a core protein by the carbohydrate xylose. The biosynthesis of GAG chains is initiated by xylosylation of the core protein followed by galactosylation by the galactosyltransferase beta 4GalT7. Some beta-D-xylosides, such as 2-naphthyl beta-D-xylopyranoside, can induce GAG synthesis by serving as acceptor substrates for beta 4GalT7 and by that also compete with the GAG synthesis on core proteins. Here we present structure-activity relationships for beta 4GalT7 and xylosides with modifications of the aromatic aglycon, using enzymatic assays, cell studies, and molecular docking simulations. The results show that the aglycons reside on the outside of the active site of the enzyme and that quite bulky aglycons are accepted. By separating the aromatic aglycon from the xylose moiety by linkers, a trend towards increased galactosylation with increased linker length is observed. The galactosylation is influenced by the identity and position of substituents in the aromatic framework, and generally, only xylosides with beta-glycosidic linkages function as good substrates for beta 4GalT7. We also show that the galactosylation ability of a xyloside is increased by replacing the anomeric oxygen with sulfur, but decreased by replacing it with carbon. Finally, we propose that reaction kinetics of galactosylation by beta 4GalT7 is dependent on subtle differences in orientation of the xylose moiety.