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

ABSTRACT: 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.

SUBMITTER: Schumann B

PROVIDER: S-EPMC7276986 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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

Schumann Benjamin B Malaker Stacy Alyse SA Wisnovsky Simon Peter SP Debets Marjoke Froukje MF Agbay Anthony John AJ Fernandez Daniel D Wagner Lauren Jan Sarbo LJS Lin Liang L Li Zhen Z Choi Junwon J Fox Douglas Michael DM Peh Jessie J Gray Melissa Anne MA Pedram Kayvon K Kohler Jennifer Jean JJ Mrksich Milan M Bertozzi Carolyn Ruth CR

Molecular cell 20200422 5

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 ...[more]

PMID: 32325029

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Dataset Information

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

Publications

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

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