Identification of proteins in excretory/secretory extracts ofEchinostoma friedi (Trematoda) from chronic and acute infections

Abstract

In the present study, we describe the investigation of Echinostoma friedi excretory/secretory products using a proteomic approach combined with the use of heterologous antibodies. We have identified 18 protein spots corresponding to ten proteins, including cytoskeletal proteins like actin, tropomyosin, and paramyosin; glycolytic enzymes like enolase, glyceraldehyde 3P dehydrogenase, and aldolase; detoxifying enzymes like GSTs; and stress proteins like heat shock protein (Hsp) 70. Among these proteins, both actin and, to a lesser extent, Hsp70, exhibited differential expression patterns between chronic and acute infections in the Echinostoma-rodent model, suggesting that these proteins may play a role in the survival within the host.

Leucine Aminopeptidase Is an Immunodominant Antigen of Fasciola hepatica Excretory and Secretory Products in Human Infections

“…Each gel spot (2D) was manually excised from the gel, washed twice with double-distilled water, and digested with sequencinggrade trypsin (Roche Molecular Biochemicals) as described previously (6,55). After digestion, 1 l of the supernatant was spotted onto a Teflon-coated plate (PerSeptive Biosystems, Framigham, MA) and allowed to air dry for 10 min at room temperature.…”

Foodborne Intestinal Flukes in Southeast Asia

“…Since recently, studies on proteomics using 2-dimensional electrophoresis (2DE) have been performed. Using 2DE, various kinds of proteins, including actin, tropomyosin, and paramyosin, glycolytic enzymes, including endolase, glyceraldehydes 3P dehydrogenase (GAPDH), and aldolase, detoxifying enzymes, in particular, glutathion-S-transferase, and the stress-related protein, Hsp 70 have been identified from E. friedi [289]. The major limitation in proteomic studies on echinostomes is the lack of sufficient genomic data.…”

Annexin A5 stimulates autophagy and inhibits endocytosis

“…Two-dimensional electrophoresis and mass spectroscopy 2D-DIGE of lysosomal membranes was carried out essentially as previously described (Bernal et al, 2006), solubilising protein samples purified as above in 7 M urea, 2 M thiourea, 4% (w/v) CHAPS, 20 mM dithiothreitol (DTT) and 2% (v/ v) Biolytes 3-10 and Bromophenol Blue (all chemicals from Bio-Rad). Samples of 100 mg protein were subjected to isoelectric focusing generated on a Bio-Rad PROTEAN H IEF Cell at 20˚C.…”