Hostname: page-component-78c5997874-xbtfd Total loading time: 0 Render date: 2024-11-15T01:56:15.817Z Has data issue: false hasContentIssue false
  • English
  • Français

Aminopeptidase-like activities in Caenorhabditis elegans and the soybean cyst nematode, Heterodera glycines

Published online by Cambridge University Press:  12 April 2024

E.P. Masler ,
E.S. Kovaleva  and
S. Sardanelli
E.P. Masler*
Affiliation:
Nematology Laboratory, United States Department of Agriculture, Agricultural Research Service, 10300 Baltimore Avenue, BARC-West, Beltsville, MD 20705-2350, USA
E.S. Kovaleva
Affiliation:
Nematology Laboratory, United States Department of Agriculture, Agricultural Research Service, 10300 Baltimore Avenue, BARC-West, Beltsville, MD 20705-2350, USA
S. Sardanelli
Affiliation:
Plant Nematology Laboratory, Department of Entomology, University of Maryland at College Park, College Park, MD 20742-5815, USA
*
*Fax: 301 504 5589 E-mail: maslere@ba.ars.usda.gov
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Aminopeptidase-like activities in crude whole body extracts of the free-living nematode Caenorhabditis elegans and the plant parasitic soybean cyst nematode Heterodera glycines were examined. General characteristics including pH optima, heat lability, and inactivation of enzyme by organic solvent were the same for the two species. All developmental stages of H. glycines exhibited activity. In older females, activity was present primarily in the eggs. Affinity for the substrate L-alanine-4-nitroanilide was the same regardless of the stage examined, and was similar for the two species (Km=2.3±0.3 mM FOR C. ELEGANS AND 2.9±0.2 Mm for H. glycines). Nearly all (>95%) of C. elegans aminopeptidase-like activity was present in the soluble fraction of the extract, while H. glycines activity was distributed between the soluble and membrane fractions. Specific activities of the soluble enzymes were highest in C. elegans and H. glycines juveniles. The C. elegans enzyme was susceptible to a number of aminopeptidase inhibitors, particularly to amastatin and leuhistin, each of which inhibited aminopeptidase-like activity more than 90% at 90 μm. In H. glycines, aminopeptidase-like activity was inhibited 39% by amastatin at 900 μm. The apparent molecular weight of the soluble C. elegans enzyme is 70–80 kDa. Some activity in H. glycines is present in the 70–80 kDa range, but most activity (80–90%) is associated with a very high molecular weight (>240 kDa) component.

Type
Research Article
Information
Journal of Helminthology , Volume 75 , Issue 3 , September 2001 , pp. 267 - 272
Copyright
Copyright © Cambridge University Press 2001

References

Atkinson, H.J., Urwin, P.E., Hansen, E. & McPherson, M.J. (1995) Designs for engineered resistance to root-parasitic nematodes. Trends in Biotechnology 13, 369–374.CrossRefGoogle Scholar
Atkinson, H.J., Lilley, C.J., Urwin, P.E. & McPherson, M.J. (1998) Engineering resistance to plant-parasitic nematodes 381–413 Perry, R.J. & Wright, D.J. (Eds) The physiology and biochemistry of free-living and plant-parasitic nematodes. Wallingford, CAB International.Google Scholar
Baset, H.A., Ford-Hutchinson, A.W. & O'Neill, G.P. (1998) Molecular cloning and functional expression of a Caenorhabditis elegans aminopeptidase structurally related to mammalian leukotriene A4 hydrolases. Journal of Biological Chemistry 273, 27978–27987.CrossRefGoogle ScholarPubMed
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
Dasgupta, D.R. & Ganguly, A.K. (1975) Isolation, purification and characterization of a trypsin-like protease from the root-knot nematode, Meloidogyne incognita . Nematologica 21, 370–384.CrossRefGoogle Scholar
Davis, R.E. & Stretton, A.O.W. (1995) Neurotransmitters of helminths. 257–287 in Marr, J.J. & Muller, M. (Eds)Biochemistry and molecular biology of parasites. New York, Academic Press.CrossRefGoogle Scholar
Gamble, H.R., Purcell, J.P. & Fetterer, R.H. (1989) Purification of a 44 kilodalton protease which mediates the ecdysis of infective Haemonchus contortus larvae. Molecular and Biochemical Parasitology 33, 49–58.CrossRefGoogle ScholarPubMed
Gimenez-Pardo, C., Vazquez-Lopez, C., de Armas-Serra, C. & Rodriguez-Caabeiro, F. (1999) Proteolytic activity in Caenorhabditis elegans: soluble and insoluble fractions. Journal of Helminthology 73, 123–127.CrossRefGoogle Scholar
Harnett, W., Houston, K.M., Tate, R., Garate, T., Apfel, H., Adam, R., Haslam, S.M., Panico, M., Paxton, T., Dell, A., Morris, H. & Brzeski, H. (1999) Molecular cloning and demonstration of an aminopeptidase activity in a filarial nematode glycoprotein. Molecular and Biochemical Parasitology 104, 11–23.CrossRefGoogle Scholar
Hotez, P., Haggerty, J., Haedon, J., Milstone, L., Gamble, H.R., Schad, G. & Richards, F. (1990) Metalloproteases of infective Ancyclostoma hookworm larvae and their possible functions in tissue invasion and ecdysis. Infection and Immunity 58, 2883–2892.CrossRefGoogle Scholar
Kim, D.G., Riggs, R.D. & Mauromoustakos, A. (1998) Variation in resistance of soybean lines to races of Heterodera glycines . Journal of Nematology 30, 184–191.Google ScholarPubMed
Knox, D.P., Redmond, D.L. & Jones, D.G. (1993) Characterization of proteinases in extracts of adult Haemonchus contortus, the ovine abomasal nematode. Parasitology 106, 395–404.CrossRefGoogle ScholarPubMed
Koritsas, V.M. & Atkinson, H.J. (1994) Proteinases of females of the phytoparasite Globodera pallida (potato cyst nematode). Parasitology 109, 357–365.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
Larminie, C.G.C. & Johnstone, I.L. (1996) Isolation and characterization of four developmentally regulated cathepsin B-like cysteine protease genes from the nematode Caenorhabditis elegans . DNA and Cell Biology 15, 75–82.CrossRefGoogle ScholarPubMed
Lilley, C.J., Urwin, P.E., McPherson, M.J. & Atkinson, H.J. (1996) Characterization of intestinally active proteinases of cyst-nematodes. Parasitology 113, 415–424.CrossRefGoogle ScholarPubMed
Lilley, C.J., Urwin, P.E., Atkinson, H.J. & McPherson, M.J. (1997) Characterization of cDNAs encoding serine proteinases from the soybean cyst nematode Heterodera glycines . Molecular and Biochemical Parasitology 89, 195–207.CrossRefGoogle ScholarPubMed
Masler, E.P., Kovaleva, E.S. & Sardanelli, S. (1999) FMRFamide-like immunoactivity in Heterodera glycines (Nemata: Tylenchida). Journal of Nematology 31, 224–231.Google ScholarPubMed
Pratt, D., Cox, G.N., Milhausen, M.J. & Boisvenue, R.J. (1990) A developmentally regulated cysteine protease gene family in Haemonchus contortus . Molecular and Biochemical Parasitology 43, 181–192.CrossRefGoogle ScholarPubMed
Rhoads, M.L. & Fetterer, R.H. (1997) Extracellular matrix: a tool for defining the extracorporeal function of parasite proteases. Parasitology Today 13, 119–122.CrossRefGoogle ScholarPubMed
Rhoads, M.L. & Fetterer, R.H. (1998) Purification and characterization of a secreted aminopeptidase from adult Ascaris suum . International Journal for Parasitology 28, 1681–1690.CrossRefGoogle ScholarPubMed
Sajid, M., Keating, C., Holden-Dye, L., Harrow, I.D. & Isaac, R.E. (1996) Metabolism of AF1 (KNEFIRF-NH2) in the nematode, Ascaris suum, by aminopeptidase, endopeptidase and deamidase enzymes. Molecular and Biochemical Parasitology 75, 159–168.CrossRefGoogle ScholarPubMed
Sajid, M., Isaac, R.E. & Harrow, I.D. (1997) Purification and properties of a membrane aminopeptidase from Ascaris suum muscle that degrades neuropeptides AF1 and AF2. Molecular and Biochemical Parasitology 89, 225–234.CrossRefGoogle ScholarPubMed
Sardanelli, S. & Kenworthy, W.J. (1997) Soil moisture control and direct seeding for bioassay of Heterodera glycines on soybean. Journal of Nematology 29(supplement), 625–634.Google ScholarPubMed
Sarkis, G.J., Kurpiewski, R., Ashcom, J.D., Jen-Jacobson, L. & Jacobson, L.A. (1988) Proteases of the nematode Caenorhabditis elegans . Archives of Biochemistry and Biophysics 261, 80–90.CrossRefGoogle ScholarPubMed
Shaw, C. (1996) Neuropeptides and their evolution. Parasitology 113, S35–S45.CrossRefGoogle ScholarPubMed
Smith, T.S., Graham, M., Munn, E.A., Newton, S.E., Knox, D.P., Coadwell, W.J., McMichael-Phillips, D., Smith, H., Smith, W.D. & Oliver, J.J. (1997) Cloning and characterization of a microsomal aminopeptidase from the intestine of the nematode Haemonchus contortus . Biochimica et Biophysica Acta 1338, 295–306.CrossRefGoogle ScholarPubMed
Taylor, A. (1993) Aminopeptidases: structure and function. FASEB Journal 7, 290–298.CrossRefGoogle ScholarPubMed
Tort, J., Brindley, P.J., Knox, D., Wolfe, K.H. & Dalton, J.P. (1999) Proteinases and associated genes of parasitic helminths. Advances in Parasitology 43, 162–266.Google ScholarPubMed
Urwin, P.E., Lilley, C.J., McPherson, M.J. & Atkinson, H.J. (1997) Characterization of two cDNAs encoding cysteine proteinases from the soybean cyst nematode Heterodera glycines . Parasitology 114, 605–613.Google ScholarPubMed
Young, L.D. (1992) Epiphytology and life cycle. 27–36 in Riggs, R.D. & Wrather, J.A. (Eds) Biology and management of the soybean cyst nematode. St Paul, American Phytopathological Society Press.Google Scholar