Protein phosphorylation networks in spargana of Spirometra erinaceieuropaei revealed by phosphoproteomic analysis

Abstract

Background Sparganosis caused by Spirometra erinaceieuropaei spargana is a zoonotic parasitic infection that has been reported in many countries, including China, Japan, Thailand and Korea, as well as European countries and the USA. The biological and clinical significance of the parasite have previously been reported. Although the genomic and transcriptomic analysis of S. erinaceieuropaei provided insightful views about the development and pathogenesis of this species, little knowledge has been acquired in terms of post-translational regulation that is essential for parasite growth, development and reproduction. Here, we performed site-specific phosphoproteomic profiling, with an aim to obtain primary information about the global phosphorylation status of spargana. Results A total of 3228 phosphopeptides and 3461 phosphorylation sites were identified in 1758 spargana proteins. The annotated phosphoproteins were involved in a variety of biological pathways, including cellular (28%), metabolic (20%) and single-organism (17%) processes. The functional enrichment of phosphopeptides by Gene Ontology analysis indicated that most spargana phosphoproteins were related to the cytoskeleton cellular compartment, signaling molecular function, and a variety of biological processes, including a molecular function regulator, guanyl-nucleotide exchange factor activity, protein kinase activities, and calcium ion binding. The highly enriched pathways of phosphorylation proteins include the phosphatidylinositol signaling system, phagosome, endocytosis, inositol phosphate metabolism, terpenoid backbone biosynthesis, and peroxisome. Domain analysis identified an EF-hand domain and pleckstrin homology domain among the key domains. Conclusions To our knowledge, this study performed the first global phosphoproteomic analysis of S. erinaceieuropaei. The dataset reported herein provides a valuable resource for future studies on the signaling pathways of this important zoonotic parasite.

A global phosphoproteomics analysis of adult Fasciola gigantica by LC–MS/MS

“…In that case, future developmental phosphoproteomics analyses supplementing with knockdown experiments are needed to confirm whether the hyper-phosphorylated glycogen synthase correlates with the development and survival of F. gigantica or not in a certain stage. On the other hand, previous studies had found that the mutations in important phosphosites might result in the termination of the phosphorylation, leading to disorders of biological processes (Liu et al 2020b ). So an additional point mutation in glycogen synthase will be used to test the essentiality of the serine or threonine phosphorylation and further explore its functional domains.…”

“…The lysate debris were eliminated by centrifugation at 12,000 g for 30 min at 4 ℃, and individual supernatants were collected and conducted to determine the protein concentration by a bicinchoninic acid (BCA) protein assay kit (Thermo Fisher Scientific, USA). Protein digestion was carried out as previously described (Liu et al 2020b ; Ren et al 2018 ), with some modifications. Briefly, 300 μg protein sample was reduced with 20 mM dithiothreitol at 56 ℃ for 30 min, followed by alkylation with 100 mM iodoacetamide at room temperature for 30 min, then dissolved into urea solution with the addition of 25 mM NH 4 HCO 3 .…”

“…However, due to the limitations in genome annotation and functional study of these kinases, there is scant knowledge of protein phosphorylation events in F. gigantica . Recent progress has been made in the global phosphoproteomics analyses of various parasites (Hirst et al 2020 ; Wang et al 2020 ; Liu et al 2020b ), which made contributions to understanding the complex biology of these organisms and developing new anti-parasitic candidates.…”

Proteomic Insights into the Biology of the Most Important Foodborne Parasites in Europe

“…Protein spots which showed immune response were characterized by MALDI-TOF/TOF-MS. The recent study on S. erinaceieuropaei sparanga described site-specific phosphoproteome, with the purpose of describing the global phosphorylation status of spargana [ 82 ]. A total of 1758 spargana proteins were identified, where 3228 phosphopeptides and 3461 phosphorylation sites were described among.…”

Molecular Characterization and Phylogenetic Analysis of Spirometra Tapeworms from Snakes in Hunan Province

“…Another reason is the high infection rate of frogs and snakes in China. A survey showed that 14.3% (31/217) and 91.7% (344/375) of frogs and snakes, respectively, were infected in Hunan Province [11,12].…”