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A comparison of the FMRFamide-like peptide proteolytic activities of preparations from two plant-parasitic nematodes (Heterodera glycines and Meloidogyne incognita): possible targets for novel control

Published online by Cambridge University Press:  06 February 2012

E.P. Masler
E.P. Masler*
Affiliation:
Nematology Laboratory, United States Department of Agriculture, Agricultural Research Service, 10300 Baltimore Avenue, BARC-West, Beltsville, MD20705-2350, USA
*
* Fax: 301-504-5062, E-mail: [email protected]
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Abstract

Proteolytic activities in extracts from the plant-parasitic nematodes Heterodera glycines and Meloidogyneincognita were examined for their abilities to digest three FRET-modified peptide substrates representing members of the large FMRFamide-like peptide (FLP) family in nematodes. Included were sequences distributed across all nematode species (KSAYMRFa and KHEYLRFa) and a sequence confined to a narrow range of plant-parasitic nematodes (KHEFVRFa). Species variations were observed among substrate affinities, reaction rates and effect of protease inhibitors. Km values for KHEYLRFa (1.48 ± 0.34 μm) and KSAYMRFa (2.13 ± 0.24 μm) in H. glycines were each lower (P< 0.05) than those for the same substrates in M. incognita (5.26 ± 1.30 μm and 3.90 ± 0.61 μm, respectively). The Km of KHEFVRFa was lower (P< 0.05) in M. incognita (5.83 ± 0.36 μm) than in H. glycines (11.01 ± 1.26 μm). Reaction rates (Vmax/min/μg) for KHEYLRFa were the same for both species, but KSAYMRFa and KHEFVRFa digestion rates were each nearly twofold higher (P< 0.05) in M. incognita than in H. glycines. Digestion of KSAYMRFa was strongly inhibited in both species by 4-(2-aminoethyl)-benzenesulfonyl-fluoride-HCl (AEBSF) and EDTA, but M. incognita was more sensitive (P< 0.05) to inhibition. AEBSF and EDTA (both at 1 mm) inhibited M. incognita activity 62.3% and 36.6% more, respectively, than H. glycines activity. Serine protease inhibition differed significantly (P< 0.05) between the two species. Maximum inhibition of M. incognita (76%) occurred at 1.85 mm AEBSF while maximum inhibition of H. glycines was 40% at 1.19 mm AEBSF.

Type
Research Papers
Information
Journal of Helminthology , Volume 87 , Issue 1 , March 2013 , pp. 71 - 77
Copyright
Copyright © Cambridge University Press 2012. This is a work of the US Government and is not subject to copyright protection in the United States

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