Sculpting the Bacterial O-Glycoproteome: Functional Analyses of Orthologous Oligosaccharyltransferases with Diverse Targeting Specificities.
Sculpting the Bacterial O-Glycoproteome: Functional Analyses of Orthologous Oligosaccharyltransferases with Diverse Targeting Specificities.
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作者：
蛋白质糖基化系统在细菌中被广泛认识，包括奈瑟氏菌属的成员。在大多数细菌物种中，支持目标蛋白选择和糖链谱系的分子机制和进化背景仍然知之甚少。广谱的O-连接蛋白糖基化发生在奈瑟氏菌属内所有与人类相关的物种群中，但对它们各自的糖蛋白谱系的了解有限。有趣的是，在淋病奈瑟菌和脑膜炎奈瑟氏菌中，IV型菌毛(TFP)定植因子的菌毛蛋白亚基PILL是糖基化的，但在深度分枝的长链奈瑟菌亚种中不是。糖酵解。为了更详细地研究这一点，我们评估了整个属的堆糖化状态，发现成堆的共生分支物种不被淋球菌PglO寡糖转移酶修饰。用淋球菌表达的整个属的PglO寡糖转移酶进行的实验表明，尽管所有酶都能够进行广谱蛋白质糖基化，但来自深分支共生体的那些酶不能支持驻留的堆糖基化。用免疫印迹和质谱仪对这些菌株进行进一步的糖蛋白质组学分析，发现其他蛋白质被其他非常相似的寡糖转移酶作为靶标。最后，我们创建了pglO等位基因嵌合体，开始定位与独特底物靶向活性相关的PglO蛋白结构域。这些发现揭示了高度相关的细菌物种的蛋白质糖基化系统中以前未被意识到的差异。我们认为，在新系列蛋白底物和寡糖转移酶中表现出的自然多样性具有很大的潜力来揭示在这些和相关的细菌蛋白糖基化系统中操作的结构-功能关系。
Protein glycosylation systems are widely recognized in bacteria, including members of the genus Neisseria. In most bacterial species, the molecular mechanisms and evolutionary contexts underpinning target protein selection and the glycan repertoire remain poorly understood. Broad-spectrum O-linked protein glycosylation occurs in all human-associated species groups within the genus Neisseria, but knowledge of their individual glycoprotein repertoires is limited. Interestingly, PilE, the pilin subunit of the type IV pilus (Tfp) colonization factor, is glycosylated in Neisseria gonorrhoeae and Neisseria meningitidis but not in the deeply branching species N. elongata subsp. glycolytica. To examine this in more detail, we assessed PilE glycosylation status across the genus and found that PilEs of commensal clade species are not modified by the gonococcal PglO oligosaccharyltransferase. Experiments using PglO oligosaccharyltransferases from across the genus expressed in N. gonorrhoeae showed that although all were capable of broad-spectrum protein glycosylation, those from a deep-branching group of commensals were unable to support resident PilE glycosylation. Further glycoproteomic analyses of these strains using immunoblotting and mass spectrometry revealed other proteins differentially targeted by otherwise remarkably similar oligosaccharyltransferases. Finally, we generated pglO allelic chimeras that begin to localize PglO protein domains associated with unique substrate targeting activities. These findings reveal previously unappreciated differences within the protein glycosylation systems of highly related bacterial species. We propose that the natural diversity manifest in the neisserial protein substrates and oligosaccharyltransferases has significant potential to inform the structure-function relationships operating in these and related bacterial protein glycosylation systems.
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影响因子：
3.2
作者：
Anonsen, Jan Haug;Vik, Ashild;Koomey, Michael
通讯作者：
Koomey, Michael
影响因子：
3.2
作者：
Borud, Bente;Aas, Finn Erik;Koomey, Michael
通讯作者：
Koomey, Michael
DOI：
10.1073/pnas.1103321108
发表时间：
2011-06-07
影响因子：
11.1
作者：
Borud, Bente;Viburiene, Raimonda;Koomey, Michael
通讯作者：
Koomey, Michael
影响因子：
4.5
作者：
Hadjineophytou, Chris;Anonsen, Jan Haug;Koomey, Michael
通讯作者：
Koomey, Michael
影响因子：
4.4
作者：
Anonsen, Jan Haug;Vik, Ashild;Koomey, Michael
通讯作者：
Koomey, Michael