In-Depth N-Glycosylation Reveals Species-Specific Modifications and Functions of the Royal Jelly Protein from Western (Apis mellifera) and Eastern Honeybees (Apis cerana)
In-Depth N-Glycosylation Reveals Species-Specific Modifications and Functions of the Royal Jelly Protein from Western (Apis mellifera) and Eastern Honeybees (Apis cerana)
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DOI：
10.1021/acs.jproteome.5b00829
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发表时间：
2015-12-01
影响因子：
4.4
通讯作者：
Li, Jianke
中科院分区：
文献类型：
--
作者：
Feng, Mao;Fang, Yu;Li, Jianke
由工蜂分泌的皇家浆（RJ）在蜜蜂生物学和人类健康中具有多种营养和防御作用。尽管据报道是糖蛋白，但RJ蛋白在不同蜜蜂物种中的糖基化特征和功能性在很大程度上是未知的。对意大利蜜蜂（Apis mellifera lingustica，Aml）和中华蜜蜂（Apis cerana cerana，Acc）的RJ蛋白进行了N-糖蛋白质组分析和功能测定。RJ产生的Aml产生80个非冗余的N-糖蛋白携带190个糖位点，其中23个新的蛋白含有35个糖位点被确定。对于Acc，首次报道了所有43种蛋白质在138个糖基化。在这两个RJ样品中观察到在糖蛋白种类、N-糖基化位点数目、糖基化基序、糖蛋白丰度水平和N-糖基化位点方面具有不同N-糖蛋白组学特征的蛋白。与Acc相比，N-糖基化主要皇家果冻蛋白2（MRJP 2）对幼虫类芽孢杆菌（P. larvae）的低抑制效率以及Aml RJ中不存在抗菌相关的糖基化阿皮霉素、透明质酸钠和围食基质的事实揭示了Aml幼虫为何对幼虫类芽孢杆菌（P. larvae）敏感的机制，幼虫类芽孢杆菌是致命的育雏病（American foulbrood，AFB）的病原体。在两个RJ样品中观察到的N-糖基化MRJP 1的抗高血压活性和在Acc中发现的比在Aml中更强的活性揭示了特异性RJ蛋白和修饰对于治疗人类高血压疾病是潜在有用的。我们的数据获得了新的理解，西方和东方蜜蜂已经进化出种特异性的糖基化策略来微调蛋白质活性，以优化分子功能，作为蜜蜂的营养素和免疫剂，并对人类的健康促进活动产生影响。这为蜜蜂和医学界有针对性地探索RJ蛋白的生物学功能提供了宝贵的资源。
Royal jelly (RJ), secreted by honeybee workers, plays diverse roles as nutrients and defense agents for honeybee biology and human health. Despite being reported to be glycoproteins, the glycosylation characterization and functionality of RJ proteins in different honeybee species are largely unknown. An in-depth N-glycoproteome analysis and functional assay of RJ produced by Apis mellifera lingustica (Aml) and Apis cerana cerana (Acc) were conducted. RJ produced by Aml yielded 80 nonredundant N-glycoproteins carrying 190 glycosites, of which 23 novel proteins harboring 35 glycosites were identified. For Acc, all 43 proteins glycosylated at 138 glycosites were reported for the first time. Proteins with distinct N-glycoproteomic characteristics in terms of glycoprotein species, number of N-glycosylated sites, glycosylation motif, abundance level of glycoproteins, and N-glycosites were observed in this two RJ samples. The fact that the low inhibitory efficiency of N-glycosylated major royal jelly protein 2 (MRJP2) against Paenibacillus larvae (P. larvae) and the absence of antibacterial related glycosylated apidaecin, hymenoptaecin, and peritrophic matrix in the Aml RJ compared to Acc reveal the mechanism for why the Aml larvae are susceptible to P. larvae, the causative agent of a fatal brood disease (American foulbrood, AFB). The observed antihypertension activity of N-glycosylated MRJP1 in two RJ samples and a stronger activity found in Acc than in Aml reveal that specific RJ protein and modification are potentially useful for the treatment of hypertensive disease for humans. Our data gain novel understanding that the western and eastern bees have evolved species-specific strategies of glycosylation to fine-tune protein activity for optimizing molecular function as nutrients and immune agents for the good of honeybee and influence on the health promoting activity for human as well. This serves as a valuable resource for the targeted probing of the biological functions of RJ proteins for honeybee and medical communities.