Bump-and-Hole Engineering Identifies Specific Substrates of Glycosyltransferases in Living Cells

Schumann, Benjamin; Malaker, Stacy Alyse; Wisnovsky, Simon Peter; Debets, Marjoke Froukje; Agbay, Anthony John; Fernandez, Daniel; Wagner, Lauren Jan Sarbo; Lin, Liang; Li, Zhen; Choi, Junwon; Fox, Douglas Michael; Peh, Jessie; Gray, Melissa Anne; Pedram, Kayvon; Kohler, Jennifer Jean; Mrksich, Milan; Bertozzi, Carolyn Ruth

doi:10.1016/j.molcel.2020.03.030

Bump-and-Hole Engineering Identifies Specific Substrates of Glycosyltransferases in Living Cells

Journal Article · Thu Jun 04 00:00:00 EDT 2020 · Molecular Cell

DOI:https://doi.org/10.1016/j.molcel.2020.03.030· OSTI ID:1637904

^[1]; Malaker, Stacy Alyse ^[2]; Wisnovsky, Simon Peter ^[2]; Debets, Marjoke Froukje ^[2]; Agbay, Anthony John ^[2]; Fernandez, Daniel ^[3]; Wagner, Lauren Jan Sarbo ^[4]; Lin, Liang ^[5]; Li, Zhen ^[6]; Choi, Junwon ^[2]; Fox, Douglas Michael ^[2]; Peh, Jessie ^[2]; Gray, Melissa Anne ^[2]; Pedram, Kayvon ^[2]; Kohler, Jennifer Jean ^[7]; Mrksich, Milan ^[5]; Bertozzi, Carolyn Ruth ^[8]

Stanford Univ., CA (United States); Francis Crick Inst., London (United Kingdom); Imperial College, London (United Kingdom)
Stanford Univ., CA (United States)
Stanford ChEM-H Macromolecular Structure Knowledge Center, Stanford, CA (United States)
Univ. of California, Berkeley, CA (United States)
Northwestern Univ., Evanston, IL (United States)
Francis Crick Inst., London (United Kingdom); Imperial College, London (United Kingdom)
Univ. of Texas Southwestern Medical Center, Dallas, TX (United States)
Stanford Univ., CA (United States); Howard Hughes Medical Inst., Chevy Chase, MD (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1637904

Journal Information:: Molecular Cell, Journal Name: Molecular Cell Journal Issue: 5 Vol. 78; ISSN 1097-2765

Publisher:: Cell Press - ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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