An alliance of gel-based and gel-free proteomic techniques displays substantial insight into the proteome of a virulent and an attenuated Histomonas meleagridis strain

ABSTRACT: The current work aimed to detect and identify significantly differentially expressed proteins between a virulent and an attenuated Histomonas meleagridis strain which were in vitro co-cultivated with Escherichia coli DH5alpha. This was done by applying 2D-DIGE and sequential window acquisition of all theoretical spectra (SWATH) MS as tools on the two well-defined strains. In the gel-based experiments, 49 identified protein spots were found to be differentially expressed, of which 37 belonged to the in vitro cultivated virulent strain and 12 to the attenuated one. The most frequently identified proteins in the virulent strain take part in cytoskeleton formation, carbohydrate metabolism and adaptation to stress. However, post-translationally modified or truncated ubiquitous cellular proteins such as actin and GAPDH were identified as upregulated in multiple gel positions. This indicated their contribution to processes not related to cytoskeleton and carbohydrate metabolism, such as fibronectin or plasminogen binding. On the other hand, proteins involved in cytoskeleton, cell division and cytoskeleton organization were frequently observed in the attenuated strain. The findings of the gel-based studies were supplemented by the gel-free SWATH MS analysis, which identified and quantified 42 significantly differentially regulated proteins. In this case proteins with peptidase activity, metabolic proteins and actin-regulating proteins were the most frequent findings in the virulent strain, while proteins involved in hydrogenosomal carbohydrate metabolism dominated the results in the attenuated one.

INSTRUMENT(S): TripleTOF 5600+, liquid chromatography separation

ORGANISM(S): Histomonas Meleagridis

TISSUE(S): Cell Culture

SUBMITTER: Ivana BILIC  

LAB HEAD: Dr. Ivana Bilic

PROVIDER: PXD009319 | Pride | 2018-11-21

REPOSITORIES: Pride