PMID:22750626

Citation	Liu, H, Zeng, H, Yao, Q, Yuan, J, Zhang, Y, Qiu, D, Yang, X, Yang, H and Liu, Z (2012) Steinernema glaseri surface enolase: Molecular cloning, biological characterization, and role in host immune suppression. Mol. Biochem. Parasitol. 185:89-98
Abstract	Entomopathogenic nematodes are widely used as biological control agents that can suppress or evade the host immune defense upon entry into insects. The surface coat of Steinernema glaseri has been shown to play important roles in defeating the host immune system. In this work, a protein fraction with antiphagocytic activity was separated by electro-elution and further analyzed by two-dimensional electrophoresis (2-DE). LC-MS/MS analysis of one protein spot from a 2-DE gel gave five peptides that were highly similar to enolases of many organisms. A 1311bp cDNA was cloned that encodes a 47kDa protein with high sequence identity to enolases from different species of nematodes. The deduced protein, Sg-ENOL, was expressed in Escherichia coli, and its glycolytic activity was demonstrated by the conversion of 2-phospho-d-glycerate to phosphoenolpyruvate. Recombinant Sg-ENOL significantly reduced the LT(50)s of Xenorhabdus poinarii and Metarhizium anisopliae when co-injected into Galleria mellonella and Locusta migratoria manilensis Meyen, respectively. Using immuno-gold transmission electron microscopy, native Sg-ENOL was confirmed to be localized to both the nematode cuticle and the surface coat. In vitro, secretion of Sg-ENOL was inducible rather than constitutive. In vivo, Sg-ENOL was detected in the host hemolymph after infection of G. mellonella with S. glaseri, indicating that Sg-ENOL was secreted into the insect hemocoel and was involved in infection. This is the first report of the cloning and characterization of a surface coat protein in an entomopathogenic nematode. Our findings provide clear evidence for an important role for a cell surface enolase in S. glaseri infection and host immune suppression.
Links	PubMed Online version:10.1016/j.molbiopara.2012.06.006
Keywords
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9BILA:B8Q027		GO:0004634: phosphopyruvate hydratase activity	ECO:0000314:		F		Figure shows separated protein bands that result from electro-eluted fractions.	complete CACAO 5255
9BILA:B8Q027	enables	GO:0004634: phosphopyruvate hydratase activity	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete

References

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