Synthesis of Aryl Azide Derivatives of UDP-GlcNAc and UDP-GalNAc and Their Use for the Affinity Labeling of Glycosyltransferases and the UDP-HexNAc Pyrophosphorylase

Identification of a UDP-GlcNAc:Skp1-Hydroxyproline GlcNAc-transferase in the Cytoplasm of Dictyostelium

“…Sepharose 6B-Thiopropyl-5-Hg-dUMP-5-Hg-dUMP was synthesized as described (19). Ethanol-precipitated 5-Hg-dUMP was washed with ethyl acetate:ethanol (1:4), dissolved in 50 ml of H 2 O, and loaded onto a 10-ml Chelex 100 resin (Bio-Rad) to remove residual Hg 2ϩ as described (20).…”

“…5-(3-Amino)allyl-UDP-GlcNAc was synthesized from 5-Hg-UDP-GlcNAc and purified over a DEAE-Sepharose Fast Flow column as described (19). The product exhibited absorbance maxima at 240 and 287 nm and a minimum at 262 nm, typical for a compound with an exocyclic double bond with a pyrimidine ring.…”

“…The photoactive aryl azide moiety of this photoaffinity probe, which has been used to label a Golgi GlcNAc-transferase (19), was linked at the same position as was the leash in the affinity resin. After exposure to UV irradiation in the presence or absence of a 23-fold excess of unlabeled UDP-GlcNAc, fractions were analyzed by SDS-PAGE and autoradiography.…”

Identification of new quorum sensing autoinducer binding partners in Pseudomonas aeruginosa using photoaffinity probes

“…This would allow the alkyl chain to function as an endogenous pseudo linker, avoiding the need for a linker moiety entirely, thereby minimizing further structural modifications to the PQS and HHQ core structures. The three most commonly used photoreactive groups are diazirines, 41 aryl azides, 50 and benzophenones. 38 Benzophenones were excluded based on their size.…”

A cell-permeable probe for the labelling of a bacterial glycosyltransferase and virulence factor

“… 3 For GTs on the other hand, only a limited number of affinity probes have been reported to date, most of which are substrate analogues based on the UDP-sugar donor. 4 This lack of chemical probes is particularly striking for bacterial GTs.…”