Horvatić Anita, Kuleš Josipa, Guillemin Nicolas, Martinković Franjo, Štimac Iva, Mrljak Vladimir, Bhide Mangesh
ERA Chair VetMedZg Project, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
Department for Parasitology and Parasitic Diseases with Clinics, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
Methods Mol Biol. 2018;1734:83-96. doi: 10.1007/978-1-4939-7604-1_9.
Constant advancements in methodology and mass spectrometry instrumentation, genome sequencing and bioinformatic tools have enabled the identification of numerous pathogen proteomes. Identifying the pathogen interacting proteins by means of high-throughput techniques is key for understanding pathogen invasion and survival mechanisms and in such a way proposing specific proteins as pharmaceutical targets. Herein we describe the methodology for the enrichment and identification of pathogen surface proteome using cell surface protein biotinylation followed by LC-MS/MS and bioinformatic analyses of such data. This strategy is to be employed for the determination of protein subcellular localization and prediction of potential pathogen interacting proteins.
方法学、质谱仪器、基因组测序和生物信息学工具的不断进步,使得众多病原体蛋白质组得以鉴定。通过高通量技术鉴定病原体相互作用蛋白,是理解病原体入侵和生存机制的关键,进而以此提出特定蛋白质作为药物靶点。在此,我们描述了利用细胞表面蛋白生物素化,随后进行液相色谱-串联质谱(LC-MS/MS)及此类数据的生物信息学分析,来富集和鉴定病原体表面蛋白质组的方法。该策略将用于确定蛋白质亚细胞定位以及预测潜在的病原体相互作用蛋白。