Characterization of a Trichinella spiralis aminopeptidase and its participation in invasion, development and fecundity.

Veterinary Research
Kai Xia GuoJing Cui

Abstract

A Trichinella spiralis aminopeptidase (TsAP) has been identified in intestinal infectious larvae (IIL) and adult worms (AW), but its biological function in the T. spiralis life cycle is unknown. The aim of this study was to characterize TsAP and ascertain its functions in the invasion, development and fecundity of T. spiralis. Recombinant TsAP (rTsAP) was expressed and purified. rTsAP has strong immunogenicity. qPCR and western blotting show that TsAP was transcribed and expressed at all T. spiralis lifecycle stages, but the expression level of TsAP mRNA and proteins at IIL and AW stages was obviously higher than those in muscle larvae (ML) and newborn larvae (NBL). The IFT results reveal that TsAP was principally located at the cuticle and the intrauterine embryos of this nematode. rTsAP had the enzymatic activity of natural aminopeptidase to hydrolyze the substrate Leu-pNA with an optimal temperature of 50 °C and optimal pH of 8.0. rTsAP promoted the larval penetration into intestinal epithelial cells, whereas anti-rTsAP antibodies suppressed the larval intrusion; the promotion and suppression was dose-dependently related to rTsAP or anti-rTsAP antibodies. TsAP protein expression level and enzymatic activity were reduced by 5...Continue Reading

References

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Datasets Mentioned

BETA
AF452239

Methods Mentioned

BETA
IEC
PCR
ELISA
confocal microscopy
IECs
transfection
confocal
co-immunoprecipitation

Software Mentioned

BOXSHADE
MEGA
CellSens
CN3D
SPSS
Clustal X
PyMOL

Related Concepts