Caught in the act: precision tools to unravel and diagnose glycoprotein misfolding

Lead Research Organisation: Imperial College London
Department Name: Dept of Chemistry


Reporters of cellular glycosylation: Engineering N-acetylglucosaminyl (GlcNAc) transferases
- One of the major types of protein glycosylation, Asn(N)-linked glycosylation, undergoes extensive maturation in the secretory pathway in an assembly line principle. Crucial bifurcation points in glycan maturation are generated through introduction of GlcNAc at defined positions (
- Dysfunctional N-glycosylation is a major cause of rare congenital disorders of glycosylation (CDGs) that are often complex and present with severe developmental/neurological symptoms.
- We have the capacity to bump-and-hole engineer the GlcNAc transferases MGAT1, MGAT2 and MGAT5 to accept bioorthogonally labeled, bumped nucleotide-sugar substrates ( to allow identification of CDG-relevant substrates and create reporter systems for glycoprotein maturation at different stages of the secretory pathway.
- Objectives:
o Engineering MGAT1, MGAT2 and/or MGAT5 to accept bumped versions of the substrate UDP-GlcNAc.
o Synthesis of bumped UDP-sugars using chemoenzymatic synthesis.
o Cellular delivery of the substrate, click/enrichment/proteomics of model glycoproteins to benchmark the approach.
o Application to profile protein trafficking by imaging (e.g. through bioorthogonal ligation with cell-permeable turn-on fluorophores).
o Establishment of a toolbox to probe protein maturation/recycling and shed light on CDGs, e.g. by


10 25 50

Studentship Projects

Project Reference Relationship Related To Start End Student Name
EP/S023518/1 30/09/2019 30/03/2028
2597991 Studentship EP/S023518/1 01/10/2021 29/09/2025 Abdul ZAFAR