Life cycle stage-resolved proteomic analysis of the excretome/secretome from Strongyloides ratti - identification of stage-specific proteases.
Creators: name: Soblik, Hanns
name: Younis, Abuelhassan Elshazly
name: Mitreva, Makedonka
name: Renard, Bernhard Y.
name: Kirchner, Marc
name: Geisinger, Frank
name: Steen, Hanno
name: Brattig, Norbert W.
Publisher: name: Robert Koch-Institut
Year: 2011
Subjects: subject: Amino Acid Sequence
subject: Gene Expression Regulation
subject: Molecular Sequence Data
subject: Female
subject: Male
subject: Animals
subject: Base Sequence
subject: Rats
subject: Protease Inhibitors/pharmacology
subject: Proteomics
subject: Culture Media/chemistry
subject: Developmental
subject: Helminth Proteins/genetics
subject: Helminth Proteins/metabolism
subject: Helminth Proteins/secretion
subject: Immune Sera/chemistry
subject: Intestines/parasitology
subject: Larva/enzymology
subject: Larva/genetics
subject: Larva/growth &amp; development
subject: Peptide Hydrolases/genetics
subject: Peptide Hydrolases/metabolism
subject: Peptide Hydrolases/secretion
subject: Protein Sorting Signals
subject: Protein Structure Tertiary
subject: Rats Wistar
subject: Real-Time Polymerase Chain Reaction
subject: Sequence Analysis Protein
subject: Serine Endopeptidases/genetics
subject: Serine Endopeptidases/metabolism
subject: Serine Endopeptidases/secretion
subject: Statistics Nonparametric
subject: Strongyloides ratti/enzymology
subject: Strongyloides ratti/genetics
subject: Strongyloides ratti/growth &amp; development
subject: Strongyloidiasis/parasitology
subject: 610 Medizin

description: A wide range of biomolecules, including proteins, are excreted and secreted from helminths and contribute to the parasite's successful establishment, survival, and reproduction in an adverse habitat. Excretory and secretory proteins (ESP) are active at the interface between parasite and host and comprise potential targets for intervention. The intestinal nematode Strongyloides spp. exhibits an exceptional developmental plasticity in its life cycle characterized by parasitic and free-living generations. We investigated ESP from infective larvae, parasitic females, and free-living stages of the rat parasite Strongyloides ratti, which is genetically very similar to the human pathogen, Strongyloides stercoralis. Proteomic analysis of ESP revealed 586 proteins, with the largest number of stage-specific ESP found in infective larvae (196), followed by parasitic females (79) and free-living stages (35). One hundred and forty proteins were identified in all studied stages, including anti-oxidative enzymes, heat shock proteins, and carbohydrate-binding proteins. The stage-selective ESP of (1) infective larvae included an astacin metalloproteinase, the L3 Nie antigen, and a fatty acid retinoid-binding protein; (2) parasitic females included a prolyl oligopeptidase (prolyl serine carboxypeptidase), small heat shock proteins, and a secreted acidic protein; (3) free-living stages included a lysozyme family member, a carbohydrate-hydrolyzing enzyme, and saponin-like protein. We verified the differential expression of selected genes encoding ESP by qRT-PCR. ELISA analysis revealed the recognition of ESP by antibodies of S. ratti-infected rats. A prolyl oligopeptidase was identified as abundant parasitic female-specific ESP, and the effect of pyrrolidine-based prolyl oligopeptidase inhibitors showed concentration- and time-dependent inhibitory effects on female motility. The characterization of stage-related ESP from Strongyloides will help to further understand the interaction of this unique intestinal nematode with its host.