Functional analysis of the Helicobacter pullorum N-linked protein glycosylation system.
Functional analysis of the Helicobacter pullorum N-linked protein glycosylation system.
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DOI：
10.1093/glycob/cwx110
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发表时间：
2018-04-01
期刊：
影响因子：
4.3
通讯作者：
Linton D
中科院分区：
文献类型：
--
作者：
Jervis AJ;Wood AG;Cain JA;Butler JA;Frost H;Lord E;Langdon R;Cordwell SJ;Wren BW;Linton D
N-连接的蛋白质糖基化系统在生命的所有三个领域的物种中运作。空肠弯曲菌的模式细菌N-连接糖基化系统是由单个染色体上存在的PGL基因编码的。这个基因簇包括负责将多糖从脂质载体转移到蛋白质的pglB寡糖转移酶。虽然所有弯曲杆菌属的基因组都含有一个PGL基因座，但在相关的螺杆菌属中，只有三个进化上有亲缘关系的物种(猪瘟杆菌、加拿大幽门螺杆菌和翼状幽门螺杆菌)可能编码N-连接蛋白糖基化系统。幽门螺杆菌可能的PGL基因分布在5个染色体上，每个基因组包括两个可能编码寡糖转移酶的pglB基因。我们已经在体外证明了鸡白羽杆菌的N-连接糖基化活性，导致五糖转移到天冬酰胺上的一个肽上(D/E)XNXS/T。在本研究中，我们鉴定了第一个N-连接的糖蛋白，命名为HgpA。在鸡白喉PgL/WBP基因插入敲除突变体的背景下产生组氨酸标记的HgpA，随后对HgpA的糖链结构进行分析，证实了单个基因产物在PglB1依赖的N-连锁蛋白糖基化途径中的作用。利用两性离子-亲水相互作用的高效液相色谱-串联质谱法从5种白喉杆菌蛋白中鉴定出6个糖位，与糖基化HgpA多克隆抗血清反应的蛋白一致。这项研究证明了鸡白疽杆菌N-连接的普通蛋白糖基化系统的功能。
N-linked protein glycosylation systems operate in species from all three domains of life. The model bacterial N-linked glycosylation system from Campylobacter jejuni is encoded by pgl genes present at a single chromosomal locus. This gene cluster includes the pglB oligosaccharyltransferase responsible for transfer of glycan from lipid carrier to protein. Although all genomes from species of the Campylobacter genus contain a pgl locus, among the related Helicobacter genus only three evolutionarily related species (H. pullorum, H. canadensis and H. winghamensis) potentially encode N-linked protein glycosylation systems. Helicobacter putative pgl genes are scattered in five chromosomal loci and include two putative oligosaccharyltransferase-encoding pglB genes per genome. We have previously demonstrated the in vitro N-linked glycosylation activity of H. pullorum resulting in transfer of a pentasaccharide to a peptide at asparagine within the sequon (D/E)XNXS/T. In this study, we identified the first H. pullorum N-linked glycoprotein, termed HgpA. Production of histidine-tagged HgpA in the background of insertional knockout mutants of H. pullorum pgl/wbp genes followed by analysis of HgpA glycan structures demonstrated the role of individual gene products in the PglB1-dependent N-linked protein glycosylation pathway. Glycopeptide purification by zwitterionic-hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry identified six glycosites from five H. pullorum proteins, which was consistent with proteins reactive with a polyclonal antiserum generated against glycosylated HgpA. This study demonstrates functioning of a H. pullorum N-linked general protein glycosylation system.
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影响因子：
5.8
作者：
Cuccui J;Terra VS;Bossé JT;Naegeli A;Abouelhadid S;Li Y;Lin CW;Vohra P;Tucker AW;Rycroft AN;Maskell DJ;Aebi M;Langford PR;Wren BW;BRaDP1T Consortium
通讯作者：
BRaDP1T Consortium
影响因子：
6.4
作者：
Iwashkiw JA;Fentabil MA;Faridmoayer A;Mills DC;Peppler M;Czibener C;Ciocchini AE;Comerci DJ;Ugalde JE;Feldman MF
通讯作者：
Feldman MF
影响因子：
2.9
作者：
Larkin A;Imperiali B
通讯作者：
Imperiali B
影响因子：
4.8
作者：
Gross, Julia;Grass, Susan;Geme, Joseph W. St., III
通讯作者：
Geme, Joseph W. St., III
影响因子：
3.6
作者：
Grass, S;Buscher, AZ;St Geme, JW
通讯作者：
St Geme, JW