From the ‡Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, Oregon.
§Department of Reproductive Health and Research, World Health Organization, Geneva, Switzerland.
Mol Cell Proteomics. 2019 Jan;18(1):127-150. doi: 10.1074/mcp.RA118.001125. Epub 2018 Oct 23.
The sexually transmitted disease gonorrhea (causative agent: ) remains an urgent public health threat globally because of its reproductive health repercussions, high incidence, widespread antimicrobial resistance (AMR), and absence of a vaccine. To mine gonorrhea antigens and enhance our understanding of gonococcal AMR at the proteome level, we performed the first large-scale proteomic profiling of a diverse panel ( = 15) of gonococcal strains, including the 2016 World Health Organization (WHO) reference strains. These strains show all existing AMR profiles - established through phenotypic characterization and reference genome publication - and are intended for quality assurance in laboratory investigations. Herein, these isolates were subjected to subcellular fractionation and labeling with tandem mass tags coupled to mass spectrometry and multi-combinatorial bioinformatics. Our analyses detected 904 and 723 common proteins in cell envelope and cytoplasmic subproteomes, respectively. We identified nine novel gonorrhea vaccine candidates. Expression and conservation of new and previously selected antigens were investigated. In addition, established gonococcal AMR determinants were evaluated for the first time using quantitative proteomics. Six new proteins, WHO_F_00238, WHO_F_00635c, WHO_F_00745, WHO_F_01139, WHO_F_01144c, and WHO_F_01126, were differentially expressed in all strains, suggesting that they represent global proteomic AMR markers, indicate a predisposition toward developing or compensating gonococcal AMR, and/or act as new antimicrobial targets. Finally, phenotypic clustering based on the isolates' defined antibiograms and common differentially expressed proteins yielded seven matching clusters between established and proteome-derived AMR signatures. Together, our investigations provide a reference proteomics data bank for gonococcal vaccine and AMR research endeavors, which enables microbiological, clinical, or epidemiological projects and enhances the utility of the WHO reference strains.
性传播疾病淋病（病原体：）仍然是全球紧迫的公共卫生威胁，因为它对生殖健康有影响、发病率高、广泛存在抗生素耐药性（AMR），而且没有疫苗。为了挖掘淋病抗原并在蛋白质组水平上增强我们对淋球菌 AMR 的理解，我们对包括 2016 年世界卫生组织（WHO）参考株在内的多样化淋球菌株（= 15）进行了首次大规模蛋白质组分析。这些菌株显示了所有现有的 AMR 图谱 - 通过表型特征描述和参考基因组发表确定 - 并旨在用于实验室研究的质量保证。在此，这些分离物进行了亚细胞分级和串联质量标签标记，结合质谱和多组合生物信息学分析。我们的分析分别在细胞膜和细胞质亚蛋白组中检测到 904 和 723 个常见蛋白质。我们鉴定了 9 种新的淋病疫苗候选物。研究了新的和以前选择的抗原的表达和保守性。此外，首次使用定量蛋白质组学评估了已建立的淋球菌 AMR 决定因素。使用定量蛋白质组学首次评估了已建立的淋球菌 AMR 决定因素。六个新的蛋白质，即 WHO_F_00238、WHO_F_00635c、WHO_F_00745、WHO_F_01139、WHO_F_01144c 和 WHO_F_01126，在所有菌株中均有差异表达，这表明它们代表全球蛋白质组 AMR 标志物，表明它们具有发展或补偿淋球菌 AMR 的倾向，并且/或者可以作为新的抗菌靶标。最后，基于分离物定义的药敏谱和常见差异表达蛋白的表型聚类产生了 7 个与已建立和蛋白质组衍生的 AMR 特征相匹配的聚类。总的来说，我们的研究为淋球菌疫苗和 AMR 研究提供了参考蛋白质组学数据库，这使微生物学、临床或流行病学项目得以开展，并提高了 WHO 参考株的效用。
