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
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
复制标题
使用合成荧光底物的新平台鉴定高尔基体中具有外切作用 β-别糖苷酶活性的新型聚糖加工酶
DOI：
10.1016/j.bmc.2014.11.023
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Toshiyuki Nishio
中科院分区：
文献类型：
--
作者：
Wataru Hakamata;Kazuki Miura;Takako Hirano;Toshiyuki Nishio
The majority of eukaryotic proteins undergo post-translational modifications (PTMs) involving the attachment of complex glycans, predominantly throughN-glycosylation andO-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.
