Protein glycosylation in Helicobacter pylori: beyond the flagellins?
Protein glycosylation in Helicobacter pylori: beyond the flagellins?
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DOI：
10.1371/journal.pone.0025722
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Creuzenet C
中科院分区：
文献类型：
--
作者：
Hopf PS;Ford RS;Zebian N;Merkx-Jacques A;Vijayakumar S;Ratnayake D;Hayworth J;Creuzenet C
假氨基酸对鞭毛蛋白的糖基化是幽门螺杆菌毒力所必需的。我们证明，在H。在幽门螺杆菌中，糖基化延伸至除鞭毛蛋白以外的蛋白质和除假氨基酸以外的糖。通过ProQ-绿宝石染色和DIG-或生物素-酰肼标记野生型H的可溶性和外膜部分，检测到与鞭毛蛋白不同的几种候选糖蛋白。pylori，表明蛋白质糖基化并不限于鞭毛蛋白。DIG-酰肼标记的蛋白质从假氨基酸生物合成途径突变体显示，一些糖蛋白的糖基化不依赖于假氨基酸糖基化途径，表明存在一种新的糖基化途径。通过离子交换色谱法富集糖蛋白候选物的级分用于通过酸水解提取糖。高效阴离子交换色谱与脉冲安培检测发现这些提取物中的特征单糖峰。然后通过LC-ESI-MS/MS鉴定单糖。光谱与糖如5，7-二乙酰氨基-3，5，7，9-四脱氧-L-甘油-L-甘露-壬酮糖酸一致（Pse 5Ac 7Ac）先前在鞭毛蛋白中描述的5-乙脒基-7-乙酰氨基-3，5，7，9-四脱氧-L-甘油-L-甘露-壬酮糖酸（Pse 5Am 7Ac）、杆菌胺衍生物和一种以前在H.幽门。这些数据为研究蛋白质糖基化对H.幽门。
Glycosylation of flagellins by pseudaminic acid is required for virulence in Helicobacter pylori. We demonstrate that, in H. pylori, glycosylation extends to proteins other than flagellins and to sugars other than pseudaminic acid. Several candidate glycoproteins distinct from the flagellins were detected via ProQ-emerald staining and DIG- or biotin- hydrazide labeling of the soluble and outer membrane fractions of wild-type H. pylori, suggesting that protein glycosylation is not limited to the flagellins. DIG-hydrazide labeling of proteins from pseudaminic acid biosynthesis pathway mutants showed that the glycosylation of some glycoproteins is not dependent on the pseudaminic acid glycosylation pathway, indicating the existence of a novel glycosylation pathway. Fractions enriched in glycoprotein candidates by ion exchange chromatography were used to extract the sugars by acid hydrolysis. High performance anion exchange chromatography with pulsed amperometric detection revealed characteristic monosaccharide peaks in these extracts. The monosaccharides were then identified by LC-ESI-MS/MS. The spectra are consistent with sugars such as 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (Pse5Ac7Ac) previously described on flagellins, 5-acetamidino-7-acetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (Pse5Am7Ac), bacillosamine derivatives and a potential legionaminic acid derivative (Leg5AmNMe7Ac) which were not previously identified in H. pylori. These data open the way to the study of the mechanism and role of protein glycosylation on protein function and virulence in H. pylori.
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DOI：
10.1073/pnas.93.25.14648
发表时间：
1996-12-10
影响因子：
11.1
作者：
Censini, S;Lange, C;Covacci, A
通讯作者：
Covacci, A
DOI：
10.1016/j.bpg.2006.04.008
发表时间：
2006-01-01
影响因子：
3.2
作者：
Forman, D.;Burley, V. J.
通讯作者：
Burley, V. J.
影响因子：
3.2
作者：
Fischer, W;Haas, R
通讯作者：
Haas, R
影响因子：
3.1
作者：
CLYNE, M;LABIGNE, A;DRUMM, B
通讯作者：
DRUMM, B
影响因子：
3.2
作者：
Baik, SC;Kim, KM;Rhee, KH
通讯作者：
Rhee, KH