@article{Li:2024:10.1101/2023.12.20.572522,
author = {Li, Z and Di, Vagno L and Chawla, H and Ni, Cheallaigh A and Critcher, M and Sammon, D and Briggs, DC and Chung, N and Chang, V and Mahoney, KE and Cioce, A and Murphy, LD and Chen, Y-H and Narimatsu, Y and Miller, RL and Willems, LI and Malaker, SA and Huang, ML and Miller, GJ and Hohenester, E and Schumann, B},
doi = {10.1101/2023.12.20.572522},
journal = {bioRxiv},
title = {Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells.},
url = {http://dx.doi.org/10.1101/2023.12.20.572522},
year = {2024}
}

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TY  - JOUR
AB  - Mammalian cells orchestrate signalling through interaction events on their surfaces. Proteoglycans are an intricate part of these interactions, carrying large glycosaminoglycan polysaccharides that recruit signalling molecules. Despite their importance in development, cancer and neurobiology, a relatively small number of proteoglycans have been identified. In addition to the complexity of glycan extension, biosynthetic redundancy in the first protein glycosylation step by two xylosyltransferase isoenzymes XT1 and XT2 complicates annotation of proteoglycans. Here, we develop a chemical genetic strategy that manipulates the glycan attachment site of cellular proteoglycans. By employing a tactic termed bump- and-hole engineering, we engineer the two isoenzymes XT1 and XT2 to specifically transfer a chemically modified xylose analogue to target proteins. The chemical modification contains a bioorthogonal tag, allowing the ability to visualise and profile target proteins modified by both transferases in mammalian cells. The versatility of our approach allows pinpointing glycosylation sites by tandem mass spectrometry, and exploiting the chemical handle to manufacture proteoglycans with defined GAG chains for cellular applications. Engineered XT enzymes permit a view into proteoglycan biology that is orthogonal to conventional techniques in biochemistry.
AU  - Li,Z
AU  - Di,Vagno L
AU  - Chawla,H
AU  - Ni,Cheallaigh A
AU  - Critcher,M
AU  - Sammon,D
AU  - Briggs,DC
AU  - Chung,N
AU  - Chang,V
AU  - Mahoney,KE
AU  - Cioce,A
AU  - Murphy,LD
AU  - Chen,Y-H
AU  - Narimatsu,Y
AU  - Miller,RL
AU  - Willems,LI
AU  - Malaker,SA
AU  - Huang,ML
AU  - Miller,GJ
AU  - Hohenester,E
AU  - Schumann,B
DO  - 10.1101/2023.12.20.572522
PY  - 2024///
TI  - Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells.
T2  - bioRxiv
UR  - http://dx.doi.org/10.1101/2023.12.20.572522
UR  - https://www.ncbi.nlm.nih.gov/pubmed/38979271
ER  - 

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