Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species
Genetic, Structural, and Antigenic Analyses of Glycan Diversity in the O-Linked Protein Glycosylation Systems of Human Neisseria Species
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DOI：
10.1128/jb.00101-10
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发表时间：
2010-06-01
影响因子：
3.2
通讯作者：
Koomey, Michael
中科院分区：
文献类型：
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作者：
Borud, Bente;Aas, Finn Erik;Koomey, Michael
细菌被膜多糖和脂多糖是天然和获得性免疫效应物的成熟配基，经常表现出结构和抗原的变异性。虽然已经在细菌病原体和共生体中发现了许多表面定位的糖蛋白，但尚不清楚选择是否以及如何影响相关的糖类结构。在这里，设计了一种系统的方法来将奈瑟氏菌的基因谱系与对人类健康和疾病重要的奈瑟氏菌的蛋白质相关糖形结构联系起来。通过控制蛋白糖基化(PGL)基因的含量，用质谱仪和与单抗的反应性来评估糖链结构，证实了蛋白相关糖链是抗原性可变的，并且在体外至少可以表达9种不同的糖型。这些研究还表明，除了淋球菌N400株外，另一株淋球菌以及脑膜炎奈瑟菌和内酰胺奈瑟菌的分离株都表现出广谱的O-连接蛋白糖基化。虽然观察到PGL基因含量、糖型表达和血清学特征之间有很强的相关性，但也有显著的例外，特别是在微观异质性水平方面。这项工作为新序列蛋白糖链的分子血清学分型和阐明PGL基因进化提供了技术平台。
Bacterial capsular polysaccharides and lipopolysaccharides are well-established ligands of innate and adaptive immune effectors and often exhibit structural and antigenic variability. Although many surface-localized glycoproteins have been identified in bacterial pathogens and symbionts, it not clear if and how selection impacts associated glycoform structure. Here, a systematic approach was devised to correlate gene repertoire with protein-associated glycoform structure in Neisseria species important to human health and disease. By manipulating the protein glycosylation (pgl) gene content and assessing the glycan structure by mass spectrometry and reactivity with monoclonal antibodies, it was established that protein-associated glycans are antigenically variable and that at least nine distinct glycoforms can be expressed in vitro. These studies also revealed that in addition to Neisseria gonorrhoeae strain N400, one other gonococcal strain and isolates of Neisseria meningitidis and Neisseria lactamica exhibit broad-spectrum O-linked protein glycosylation. Although a strong correlation between pgl gene content, glycoform expression, and serological profile was observed, there were significant exceptions, particularly with regard to levels of microheterogeneity. This work provides a technological platform for molecular serotyping of neisserial protein glycans and for elucidating pgl gene evolution.