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

• DOI: 10.1016/j.molcel.2020.03.030
• Publication Date: 2020-04-22T14:50:09Z
• Authors: Benjamin Schumann; Stacy Alyse Malaker; Simon Peter Wisno...; Marjoke Froukje D...; Anthony John Agbay; Daniel Fernandez; Lauren Jan Sarbo ...; Liang Lin; Zhen Li; Junwon Choi; Douglas Michael Fox; Jessie Peh; Melissa Anne Gray; Kayvon Pedram; Jennifer Jean Kohler; Milan Mrksich; Carolyn Ruth Bert...
• Keywords: bioorthogonal; 570; Biochemistry & Molecular Biology; Glycosylation; Protein Engineering; glycosyltransferase; VALIDATION; Article; O-GLYCOSYLATION; chemical proteomics; mucin; CHEMISTRY; Polysaccharides; COFACTOR; GLYCOPROTEOME; Humans; TRANSCRIPTION; 11 Medical and Health Sciences; O-glycosylation; Science & Technology; Cell Membrane; Glycosyltransferases; Proteins; Cell Biology; Hep G2 Cells; 06 Biological Sciences; GLCNAC-MODIFIED PROTEINS; 620; Biosynthetic Pathways; UDP-GALNAC; HEK293 Cells; DISCOVERY; Polypeptide N-acetylgalactosaminyltransferase; N-Acetylgalactosaminyltransferases; METHYLTRANSFERASE; isoenzyme; K562 Cells; Life Sciences & Biomedicine; Developmental Biology

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 regulation is currently elusive. Here, we engineer cells tag glycans with editable chemical functionalities while providing information biosynthesis, physiological context, fine structure. We introduce a non-natural substrate pathway use engineered glycosyltransferases incorporate chemically tagged sugars into cell surface glycome cell. apply strategy particularly redundant yet human glycosyltransferase family, polypeptide N-acetylgalactosaminyl transferases. This approach bestows gain-of-chemical-functionality modification cells, where products individual can be selectively characterized or manipulated understand contribution major processes.