RRC ID 58963
Author Hakamata W, Miura K, Hirano T, Nishio T.
Title Identification of a novel glycan processing enzyme with exo-acting β-allosidase activity in the Golgi apparatus using a new platform for the synthesis of fluorescent substrates.
Journal Bioorg Med Chem
Abstract The majority of eukaryotic proteins undergo post-translational modifications (PTMs) involving the attachment of complex glycans, predominantly through N-glycosylation and O-glycosylation. PTMs play important roles in virtually all cellular processes, and aberrant regulation of protein glycosylation and glycan processing has been implicated in various diseases. However, glycan processing on proteins in various cellular contexts has not been visualized. We had previously developed a quinone methide cleavage (QMC) platform for enhanced substrate design. This platform was applied here to screen for novel glycan-processing enzymes. We designed and synthesized fluorescent substrates with β-allopyranoside residues using the QMC platform. When applied in cell-based assays, the fluorescent substrates allowed rapid and clear visualization of β-allosidase activity in the Golgi apparatus of human cultured cells. The QMC platform will likely find broad applications in visualizing the activities of glycan processing enzymes in living cells and in studying PTMs.
Volume 23(1)
Pages 73-9
Published 2015-1-1
DOI 10.1016/j.bmc.2014.11.023
PII S0968-0896(14)00810-4
PMID 25497961
MeSH Cell Line Fluorescent Dyes / chemistry* Glycosides / chemistry Glycosides / metabolism* Glycosylation Golgi Apparatus / enzymology* Golgi Apparatus / metabolism Humans Polysaccharides / chemistry Polysaccharides / metabolism* Protein Processing, Post-Translational
IF 3.073
Times Cited 4
Resource
Human and Animal Cells HeLa(RCB0007) SK-N-SH(RCB0426) HT1080(RCB1956)