Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria.
Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria.
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由深海通风蛋白细菌编码的N-链接寡核酸转移酶的功能分析。
DOI：
10.1093/glycob/cwv111
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发表时间：
2016-04
期刊：
影响因子：
4.3
通讯作者：
Wren BW
中科院分区：
文献类型：
--
作者：
Mills DC;Jervis AJ;Abouelhadid S;Yates LE;Cuccui J;Linton D;Wren BW
细菌N-连接寡糖基转移酶（OTase酶）将脂连接聚糖转移至周质中的选定蛋白质，并首次在肠道病原体空肠弯曲杆菌（细菌的ε-变形菌亚门的成员）中描述。最近，已经描述了来自其他ε-变形菌弯曲杆菌和螺杆菌物种以及δ-变形菌脱硫脱硫弧菌的直向同源物，这表明细菌的这两个亚群可能是进一步的N-连接蛋白糖基化系统的来源。深海喷口栖息地的ε-和δ-变形菌的全基因组测序，这些亚类的丰富物种来源，揭示了推定的ORF编码OTase酶和相关的邻近糖基转移酶类似于C。空肠N-连接糖基化位点。我们在大肠杆菌中表达了来自深海喷口物种Nitratiruptor tergarcus、Sulfurovum lithotrophicum和Deferribacter sulfuricans的推定OTase ORF，并表明它们能够在功能上补充C.空肠OTase、CjPglB .这些酶被证明具有松弛的聚糖特异性，转移不同的聚糖结构，并表现出不同的糖基化序列子特异性。一个宽容的D。脱硫菌受体蛋白，我们提供的证据表明，N-连接的聚糖合成的N。tergarcus和S.石养型在还原端含有乙酰化糖。这项工作表明，深海喷口细菌编码功能性N-糖基化机制，是生物技术上重要的OTase酶的潜在来源。
Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed they were able to functionally complement the C. jejuni OTase, CjPglB . The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. Additionally a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesised by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes.