Biochemical evidence for an alternate pathway in N-linked glycoprotein biosynthesis.

Biochemical evidence for an alternate pathway in N-linked glycoprotein biosynthesis.
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DOI:
10.1038/nchembio.1249
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发表时间:
2013-06
影响因子:
14.8
通讯作者:
Imperiali, Barbara
Imperiali, Barbara
中科院分区:
生物学1区
文献类型:
--
作者:
Larkin, Angelyn;Chang, Michelle M.;Whitworth, Garrett E.;Imperiali, Barbara

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天冬酰胺连接的糖基化是一种复杂的蛋白质修饰,在生命的所有三个领域中都是保守的。在此,我们报告了产甲烷古菌Methanococcus pecae的N-连接糖基化的体外分析。使用一套合成和半合成底物,我们表明AglK启动M中的N-连接糖基化。通过形成α-连接的一磷酸二氢钠N-乙酰葡糖胺(Dol-P-GlcNAc)来分解,这与真核生物和细菌中的聚异戊二烯基二磷酸中间体形成对比。有趣的是,AglK表现出与真核生物中的长链磷酸β-葡糖基转移酶(包括Alg 5)的高度序列同源性,表明共同的进化起源。前两种酶AglK和AglC的联合作用提供了α-连接的Dol-P-聚糖,其充当古细菌寡糖基转移酶AglB的感受态底物。这些研究提供了第一个生化证据,揭示了尽管整体途径明显相似,但实际上有两种一般策略来实现跨生命领域的N-连接糖蛋白。
Asparagine-linked glycosylation is a complex protein modification conserved among all three domains of life. Herein we report the in vitro analysis of N-linked glycosylation from the methanogenic archaeon Methanococcus voltae. Using a suite of synthetic and semisynthetic substrates, we show that AglK initiates N-linked glycosylation in M. voltae through the formation of α-linked dolichyl monophosphate N-acetylglucosamine (Dol-P-GlcNAc), which contrasts with the polyprenyl-diphosphate intermediates that feature in both eukaryotes and bacteria. Intriguingly, AglK exhibits high sequence homology to dolichyl-phosphate β-glucosyltransferases, including Alg5 in eukaryotes, suggesting a common evolutionary origin. The combined action of the first two enzymes, AglK and AglC, afforded an α-linked Dol-P-glycan that serves as a competent substrate for the archaeal oligosaccharyl transferase AglB. These studies provide the first biochemical evidence revealing that despite the apparent similarity of the overall pathways, there are actually two general strategies to achieve N-linked glycoproteins across the domains of life.
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