TY - JOUR
T1 - Bump-and-Hole Engineering Identifies Specific Substrates of Glycosyltransferases in Living Cells
AU - Schumann, Benjamin
AU - Malaker, Stacy Alyse
AU - Wisnovsky, Simon Peter
AU - Debets, Marjoke Froukje
AU - Agbay, Anthony John
AU - Fernandez, Daniel
AU - Wagner, Lauren Jan Sarbo
AU - Lin, Liang
AU - Li, Zhen
AU - Choi, Junwon
AU - Fox, Douglas Michael
AU - Peh, Jessie
AU - Gray, Melissa Anne
AU - Pedram, Kayvon
AU - Kohler, Jennifer Jean
AU - Mrksich, Milan
AU - Bertozzi, Carolyn Ruth
N1 - Publisher Copyright:
© 2020 The Author(s)
PY - 2020/6/4
Y1 - 2020/6/4
N2 - 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.
AB - 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.
KW - O-glycosylation
KW - bioorthogonal
KW - chemical proteomics
KW - glycosyltransferase
KW - isoenzyme
KW - mucin
UR - http://www.scopus.com/inward/record.url?scp=85085548431&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85085548431&partnerID=8YFLogxK
U2 - 10.1016/j.molcel.2020.03.030
DO - 10.1016/j.molcel.2020.03.030
M3 - Article
C2 - 32325029
AN - SCOPUS:85085548431
SN - 1097-2765
VL - 78
SP - 824-834.e15
JO - Molecular cell
JF - Molecular cell
IS - 5
ER -