Neely Benjamin A, Janech Michael G, Fenton M Brock, Simmons Nancy B, Bland Alison M, Becker Daniel J
Chemical Sciences Division, National Institute of Standards and Technology, NIST Charleston, Charleston, South Carolina 29412, United States.
Hollings Marine Laboratory, Charleston, South Carolina 29412, United States.
J Proteome Res. 2021 May 7;20(5):2547-2559. doi: 10.1021/acs.jproteome.0c00995. Epub 2021 Apr 10.
Bats are increasingly studied as model systems for longevity and as natural hosts for some virulent viruses. Yet the ability to characterize immune mechanisms of viral tolerance and to quantify infection dynamics in wild bats is often limited by small sample volumes and few species-specific reagents. Here, we demonstrate how proteomics can overcome these limitations by using data-independent acquisition-based shotgun proteomics to survey the serum proteome of 17 vampire bats () from Belize. Using just 2 μL of sample and relatively short separations of undepleted serum digests, we identified 361 proteins across 5 orders of magnitude. Levels of immunological proteins in vampire bat serum were then compared to human plasma via published databases. Of particular interest were antiviral and antibacterial components, circulating 20S proteasome complex and proteins involved in redox activity. Lastly, we used known virus proteomes to putatively identify Rh186 from and ORF1a from Middle East respiratory syndrome-related coronavirus, indicating that mass spectrometry-based techniques show promise for pathogen detection. Overall, these results can be used to design targeted mass-spectrometry assays to quantify immunological markers and detect pathogens. More broadly, our findings also highlight the application of proteomics in advancing wildlife immunology and pathogen surveillance.
蝙蝠作为长寿的模型系统以及某些烈性病毒的天然宿主,正受到越来越多的研究。然而,由于样本量小且物种特异性试剂少,表征蝙蝠病毒耐受性免疫机制和量化野生蝙蝠感染动态的能力常常受到限制。在此,我们展示了蛋白质组学如何通过基于数据非依赖采集的鸟枪法蛋白质组学克服这些限制,以检测来自伯利兹的17只吸血蝙蝠的血清蛋白质组。仅使用2微升样本和对未耗尽血清消化物进行相对较短的分离,我们就在5个数量级上鉴定出了361种蛋白质。然后通过已发表的数据库将吸血蝙蝠血清中的免疫蛋白水平与人类血浆进行比较。特别令人感兴趣的是抗病毒和抗菌成分、循环20S蛋白酶体复合物以及参与氧化还原活性的蛋白质。最后,我们使用已知的病毒蛋白质组来推定鉴定来自[具体病毒名称未给出]的Rh186和中东呼吸综合征相关冠状病毒的ORF1a,这表明基于质谱的技术在病原体检测方面具有前景。总体而言,这些结果可用于设计靶向质谱分析来量化免疫标记物和检测病原体。更广泛地说,我们的研究结果还突出了蛋白质组学在推进野生动物免疫学和病原体监测方面的应用。