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Characterization of intestinally active proteinases of cystnematodes

Published online by Cambridge University Press:  06 April 2009

C. J. Lilley ,
P. E. Urwin ,
M. J. McPherson  and
H. J. Atkinson
C. J. Lilley*
Affiliation:
Centre for Plant Biochemistry and Biotechnology, University of Leeds, Leeds LS2 9JT, UK
P. E. Urwin
Affiliation:
Centre for Plant Biochemistry and Biotechnology, University of Leeds, Leeds LS2 9JT, UK
M. J. McPherson
Affiliation:
Centre for Plant Biochemistry and Biotechnology, University of Leeds, Leeds LS2 9JT, UK
H. J. Atkinson
Affiliation:
Centre for Plant Biochemistry and Biotechnology, University of Leeds, Leeds LS2 9JT, UK
*
*Corresponding author. Tel: 0113 233 2863. Fax: 0113 233 3144.
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Summary

Cryostat sections of juvenile and adult female stages of the soybean cyst-nematode, Heterodera glycines, were incubated with 4 different naphthylamide-linked peptide substrates to localize and characterize proteinase activity within the animal. Detected activity was restricted to the intestine and 2 distinct classes of proteinase were identified on the basis of substrate specificity and sensitivity to plant proteinase inhibitors. A cathepsin L-like cysteine proteinase activity capable of hydrolysing the synthetic substrates Z-Ala-Arg-Arg-MNA and Z-Phe-Arg-MNA but not Z-Arg-Arg-MNA or L-Arg-NA was inhibited by an engineered variant of a cysteine proteinase inhibitor from rice (Oc-IδD86). The cleavage of Z-Phe-Arg-MNA was sensitive to inhibition by a combination of Oc-IδD86 and cowpea trypsin inhibitor (CpTI). Degenerate oligonucleotide primers were used to amplify fragments of cysteine proteinase genes from 2 cyst-nematodes, H. glycines and Globodera pallida. Comparison of the H. glycines fragment with known genes established highest homology to cathepsin L-like genes. In contrast, the amplified G. pallida fragment displayed greatest homology to cathepsin B-like genes from Caenorhabditis elegans.

Keywords

Heterodera glycinescyst-nematodesproteinases
Type
Research Article
Information
Parasitology , Volume 113 , Issue 4 , October 1996 , pp. 415 - 424
Copyright
Copyright © Cambridge University Press 1996

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