Bump-and-hole engineering identifies specific substrates of glycosyltransferases in living cells

Published: 20 March 2020| Version 2 | DOI: 10.17632/nh4vww6hxj.2
Benjamin Schumann


The glycosyltransferases GAlNAc-T1 and GalNAc-T2 were engineered to accommodate chemically modified UDP-GalNAc analogs. Wild type and engienered (Bump-and-hole, "BH") enzymes were stably expressed in HepG2 cells with a VSV-G epitope tag under a doxycycline-inducible promoter. Cells were treated with Dox, fixed, permeabilized and stained for VSV-G, the Golgi marker giantin and DAPI.