10779/crick.12472733.v1
Benjamin Schumann
Benjamin
Schumann
Stacy Alyse Malaker
Stacy Alyse
Malaker
Simon Peter Wisnovsky
Simon Peter
Wisnovsky
Marjoke Froukje Debets
Marjoke Froukje
Debets
Anthony John Agbay
Anthony John
Agbay
Daniel Fernandez
Daniel
Fernandez
Lauren Jan Sarbo Wagner
Lauren Jan Sarbo
Wagner
Liang Lin
Liang
Lin
Zhen Li
Zhen
Li
Junwon Choi
Junwon
Choi
Douglas Michael Fox
Douglas Michael
Fox
Jessie Peh
Jessie
Peh
Melissa Anne Gray
Melissa Anne
Gray
Kayvon Pedram
Kayvon
Pedram
Jennifer Jean Kohler
Jennifer Jean
Kohler
Milan Mrksich
Milan
Mrksich
Carolyn Ruth Bertozzi
Carolyn Ruth
Bertozzi
Bump-and-hole engineering identifies specific substrates of glycosyltransferases in living cells.
The Francis Crick Institute
2020
O-glycosylation
bioorthogonal
chemical proteomics
glycosyltransferase
isoenzyme
mucin
Schumann FC001749
CS-ack
06 Biological Sciences
11 Medical and Health Sciences
Developmental Biology
2020-06-25 14:04:16
Journal contribution
https://crick.figshare.com/articles/journal_contribution/Bump-and-hole_engineering_identifies_specific_substrates_of_glycosyltransferases_in_living_cells_/12472733
Studying posttranslational modifications classically relies on experimental strategies that oversimplify the complex biosynthetic machineries of living cells. Protein glycosylation contributes to essential biological processes, but correlating glycan structure, underlying protein, and disease-relevant biosynthetic regulation is currently elusive. Here, we engineer living cells to tag glycans with editable chemical functionalities while providing information on biosynthesis, physiological context, and glycan fine structure. We introduce a non-natural substrate biosynthetic pathway and use engineered glycosyltransferases to incorporate chemically tagged sugars into the cell surface glycome of the living cell. We apply the strategy to a particularly redundant yet disease-relevant human glycosyltransferase family, the polypeptide N-acetylgalactosaminyl transferases. This approach bestows a gain-of-chemical-functionality modification on cells, where the products of individual glycosyltransferases can be selectively characterized or manipulated to understand glycan contribution to major physiological processes.