Mitchell Natalie M, Dasari Surendra, Grys Thomas E, Lake Douglas F
School of Life Sciences, Mayo Clinic Collaborative Research Building, Arizona State University, Scottsdale, AZ 85259, USA.
Department of Health Sciences Research, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
J Fungi (Basel). 2020 Dec 14;6(4):365. doi: 10.3390/jof6040365.
Laser capture microdissection (LCM) coupled to label-free quantitative mass spectrometry is a viable strategy to identify biomarkers from infected tissues. In this study, LCM was employed to take a "snapshot" of proteins produced in vivo during spp. infection in human lungs. Proteomic analysis of LCM lung sections revealed hundreds of hosts and Coccidioidal proteins. Twenty-seven highly abundant spp. proteins were identified which do not share significant sequence orthology with human proteins. Three of the 27 Coccidioidal proteins are also potential -specific biomarkers, as they also do not share sequence homology to any other pathogenic fungus or microbe. Gene ontology analysis of the 27 biomarker candidate proteins revealed enriched hydrolase activity and increased purine and carbohydrate metabolism functions. Finally, we provide proteomic evidence that all 27 biomarker candidates are produced by the fungus when grown in vitro in a media- and growth-phase dependent manner.
激光捕获显微切割（LCM）结合无标记定量质谱分析是从感染组织中鉴定生物标志物的一种可行策略。在本研究中，采用LCM对人类肺部感染 菌期间体内产生的蛋白质进行“快照”。对LCM肺切片的蛋白质组分析揭示了数百种宿主和球孢子菌蛋白质。鉴定出27种高度丰富的球孢子菌蛋白质，它们与人类蛋白质没有明显的序列同源性。这27种球孢子菌蛋白质中的三种也是潜在的特异性生物标志物，因为它们与任何其他致病真菌或微生物也没有序列同源性。对这27种候选生物标志物蛋白质的基因本体分析显示水解酶活性增强，嘌呤和碳水化合物代谢功能增加。最后，我们提供了蛋白质组学证据，证明所有27种候选生物标志物在体外培养基中生长时由真菌以培养基和生长阶段依赖性方式产生。
