Hostname: page-component-586b7cd67f-2brh9 Total loading time: 0 Render date: 2024-11-27T02:34:19.776Z Has data issue: false hasContentIssue false
  • English
  • Français

In vitro comparison of protease activities in preparations from free-living (Panagrellus redivivus) and plant-parasitic (Meloidogyne incognita) nematodes using FMRFa and FMRFa-like peptides as substrates

Published online by Cambridge University Press:  25 March 2010

E.P. Masler
E.P. Masler*
Affiliation:
Nematology Laboratory, United States Department of Agriculture, Agricultural Research Service, 10300 Baltimore Avenue, BARC-West, Beltsville, MD 20705-2350, USA
*
*Fax: 301-504-5589 E-mail: edward.masler@ars.usda.gov
Get access Rights & Permissions [Opens in a new window]

Abstract

Extracts prepared from the microbivorous nematode Panagrellus redivivus and the plant-parasitic nematode Meloidogyne incognita were used to provide general protease activities for peptide substrate screening and species comparisons. Each extract was evaluated for its ability to degrade a broad range of nematode FMRFamide-like peptides (FLPs), key regulatory messengers governing nematode growth and development. Clear quantitative differences between the two extracts were observed using FMRFamide as a substrate. Extract potency assessed at EC50 (μg/μ l extract protein for 50% substrate digestion) was 1.8-fold greater for P. redivivus than for M. incognita, and potency assessed at EC90 was 2.5-fold greater. An overall potency difference was also present when screening the digestion of 17 nematode FLPs, but it was not universal. The mean percentage digestion of eight of the 17 FLPs was greater (P < 0.02) with P. redivivus extract (76.3 ± 8.2) than with M. incognita extract (38.1 ± 8.7), but the means for the other nine FLPs were not different. Three FLPs (KPSFVRFa, AQTFVRFa, RNKFEFIRFa) were degraded extensively by the extracts of both species, and two FLPs (SAPYDPNFLRFa, SAEPFGTMRFa) were degraded 2.9-fold and 5.3-fold greater, respectively, with M. incognita extract than with P. redivivus extract. The ability of each extract to degrade FMRFa and KSAYMRFa was significantly reduced by using peptide analogues containing single d-amino acid substitutions, and the substitution effects were positional. Both FMRFa and KSAYMRFa were competitive substrates for aminopeptidases in each extract, but only the competitive ability of FMRFa was reduced by d-amino acid substitution. The variety and complexity of nematode FLP degradation by preparations representing phylogenetically and developmentally different nematode sources are discussed.

Type
Research Papers
Information
Journal of Helminthology , Volume 84 , Issue 4 , December 2010 , pp. 425 - 433
Copyright
Copyright © Cambridge University Press 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Chitwood, D.J., Lusby, W.R., Thompson, M.J., Kochansky, J.P. & Howarth, O.W. (1995) The glycosylceramides of the nematode Caenorhabditis elegans contain an unusual, branched chain sphingoid base. Lipids 30, 567–573.CrossRefGoogle ScholarPubMed
Cohen, M., Reale, V., Olofsson, B., Knights, A., Evans, P. & de Bono, M. (2009) Coordinated regulation of foraging and metabolism in C. elegans by RFamide neuropeptide signaling. Cell Metabolism 9, 375–385.CrossRefGoogle Scholar
Craig, H., Isaac, R.E. & Brooks, D.R. (2007) Unraveling the moulting degradome: new opportunities for chemotherapy? Trends in Parasitology 23, 248–253.CrossRefGoogle ScholarPubMed
Geiss-Friedlander, R., Parmentier, N., Moeller, U., Urlaub, H., Van den Eynde, B.J. & Melchior, F. (2009) The cytoplasmic peptidase DPP9 is rate-limiting for degradation of proline-containing peptides. Journal of Biological Chemistry 284, 27211–27219.CrossRefGoogle ScholarPubMed
Greenwood, K., Williams, T. & Geary, T. (2005) Nematode neuropeptide receptors and their development as anthelmintic screens. Parasitology 131, S169–S177.CrossRefGoogle ScholarPubMed
Hong, S.Y., Oh, J.E. & Lee, K.-H. (1999) Effect of d-amino acid substitution on the stability, the secondary structure, and the activity of membrane-active peptide. Biochemical Pharmacology 58, 1775–1780.CrossRefGoogle ScholarPubMed
Husson, S.J., Mertens, I., Janssen, T., Lindemans, M. & Schoofs, L. (2007) Neuropeptidergic signaling in the nematode Caenorhabditis elegans. Progress in Neurobiology 82, 33–55.Google Scholar
Kimber, M.J. & Fleming, C.C. (2005) Neuromuscular function in plant parasitic nematodes: a target for novel control strategies? Parasitology 131, S129–S142.CrossRefGoogle ScholarPubMed
Kimber, M.J., McKinney, S., McMaster, S., Day, T.A., Fleming, C.C. & Maule, A.G. (2007) flp gene disruption in a parasitic nematode reveals motor dysfunction and unusual neuronal sensitivity to RNA interference. FASEB Journal 21, 1233–1243.CrossRefGoogle Scholar
Kubiak, T.M., Maule, A.G., Marks, N.J., Martin, R.A. & Wiest, J.R. (1996) Importance of the proline residue to the functional activity and metabolic stability of the nematode FMRFamide-related peptide, KPNFIRFamide (PF4). Peptides 17, 1267–1277.CrossRefGoogle Scholar
Li, C. (2005) The ever-expanding neuropeptide gene families in the nematode Caenorhabditis elegans. Parasitology 131, S109–S127.Google Scholar
Liu, T., Kim, K., Li, C. & Barr, M.M. (2007) FMRFamide-like peptides and mechanosensory touch receptor neurons regulate male sexual turning behavior in Caenorhabditis elegans. Journal of Neuroscience 27, 7174–7182.CrossRefGoogle ScholarPubMed
Lopez-Otin, C. & Overall, C.M. (2002) Protease degradomics: a new challenge for proteomics. Nature Reviews, Molecular Cell Biology 3, 509–519.CrossRefGoogle ScholarPubMed
Masler, E.P. (2007) Characterization of aminopeptidase in the free-living nematode Panagrellus redivivus: subcellular distribution and possible role in neuropeptide metabolism. Journal of Nematology 39, 153–160.Google Scholar
Masler, E.P. (2008a) Invertebrate neuropeptides. pp. 257–271in Meyers, R.A. (Ed.) Neurobiology. From molecular basis to disease. Weinheim, Wiley-VCH.Google Scholar
Masler, E.P. (2008b) Digestion of invertebrate neuropeptides by preparations from the free-living nematode Panagrellus redivivus. Journal of Helminthology 82, 279–285.CrossRefGoogle ScholarPubMed
Masler, E.P., Kovaleva, E.S. & Sardanelli, S. (2001) Aminopeptidase-like activities in Caenorhabditis elegans and the soybean cyst nematode, Heterodera glycines. Journal of Helminthology 75, 267–272.Google ScholarPubMed
Maule, A.G., Mousley, A., Marks, N.J., Day, T.A., Thompson, D.P., Geary, T.G. & Halton, D.W. (2002) Neuropeptide signaling system – potential drug targets for parasite control. Current Topics in Medicinal Chemistry 2, 733–758.Google Scholar
McVeigh, P., Leech, S., Mair, G.R., Marks, N.J., Geary, T.G. & Maule, A.G. (2005) Analysis of FMRFamide-like peptide (FLP) diversity in phylum Nematoda. International Journal for Parasitology 35, 1043–1060.CrossRefGoogle ScholarPubMed
McVeigh, P., Geary, T.G., Marks, N.J. & Maule, A.G. (2006) The FLP-side of nematodes. Trends in Parasitology 22, 385–396.CrossRefGoogle ScholarPubMed
Moffett, C.L., Beckett, A.M., Mousley, A., Geary, T.G., Marks, N.J., Halton, D.W., Thompson, D.P. & Maule, A.G. (2003) The ovijector of Ascaris suum: multiple response types revealed by Caenorhabditis elegans FMRFamide-related peptides. International Journal for Parasitology 33, 859–876.CrossRefGoogle ScholarPubMed
Papaioannou, S., Marsden, D., Franks, C.J., Walker, R.J. & Holden-Dye, L. (2005) Role of a FMRFamide-like family of neuropeptides in the pharyngeal nervous system of Caenorhabditis elegans. Journal of Neurobiology 65, 304–319.CrossRefGoogle ScholarPubMed
Price, D.A. & Greenberg, M.J. (1977a) Purification and characterization of a cardioexcitatory neuropeptide from the central ganglia of a bivalve mollusc. Preparative Biochemistry 7, 261–281.Google Scholar
Price, D.A. & Greenberg, M.J. (1977b) Structure of a molluscan cardioexcitatory neuropeptide. Science 197, 670–671.CrossRefGoogle ScholarPubMed
Quesada, V., Ordonez, G.R., Sanchez, L.M., Puente, X.S. & Lopez-Otin, C. (2009) The degradome database: mammalian proteases and diseases of proteolysis. Nucleic Acids Research 37(Database issue), D239–D243.CrossRefGoogle ScholarPubMed
Rogers, C., Reale, V., Kim, K., Chatwin, H., Li, C., Evans, P. & de Bono, M. (2003) Inhibition of Caenorhabditis elegans social feeding by FMRFamide-related peptide activation of NPR-1. Nature Neuroscience 6, 1178–1185.Google Scholar
Sardanelli, S. & Kenworthy, W.J. (1997) Soil moisture control and direct seeding for bioassay of Heterodera glycines on soybean. Journal of Nematology 29, 625–634.Google Scholar