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Czasopismo

1994 | 39 | 1 |

Tytuł artykułu

Ostertagia circumcincta: surface and cuticular proteins and glycoproteins isolated by biotin-streptavidin affinity chromatography

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Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
Surface proteins and glycoproteins of ex- sheathed L3 (XL3), L4 and adults were identified by labelling live worms with NHS-SS-biotin which labels primary amine groups and biotin hydrazide (BH) which introduces biotin into sugar residues. After labelling and extensive washing, worms were homogenised in low-osmotic-strength buffer (TBS) and subjected to sequential extractions with SDS and SDS + 2-Me containing buffers. Labelled proteins released from the worms by these procedures were analysed by SDS-PAGE followed by Western blotting and probing with AP-streptavidin. Western blot analysis of biotinylated cuticular proteins from parasitic stages of O. circumcincta revealed stage specific differences in the labelling patterns of the cuticular proteins and glycoproteins. Developmental differences in cuticular components were apparent in the variety of MW of the components seen in Western blots although some components identified in Western blots appeared to be conserved (78 and 45 kDa). Two to six polypeptides were recognised in XL3 surface extracts by serum from sheep vaccinated with adult nematode somatic proteins. The surface of fourth stage larvae showed less cross-reactivity. Interestingly, the molecular weights of polypeptides recognised by anti-adult serum in streptavidin affinity isolated XL3 cuticular proteins were different from those recognised in adult cuticular proteins, which suggested that the cross-reacting epitopes were located on different proteins.

Wydawca

-

Czasopismo

Rocznik

Tom

39

Numer

1

Opis fizyczny

p.46-52,fig.

Twórcy

  • Department of Parasitology, Veterinary Faculty, Warsaw Agricultural University, Grochowska 272, 03-849 Warsaw, Poland

Bibliografia

  • Betschart B., Wyss K. 1990. Analysis of the cuticular collagens of Ascaris suum. Acta Tropica, 47, 297-305.
  • Cox G. N. 1990. Molecular biology of the cuticle collagen gene families of Caenorhabditis elegans and Haemonchus contortus. Acta Tropica, 47, 269-291.
  • Cox G. N. 1992. Molecular and biochemical aspects of nematode collagens. Journal of Parasitology, 78, 1-15.
  • Cox G. N., Kusch M., Edgar R. S. 1981. Cuticle of Caenorhabditis elegans: its isolation and partial characterisation. Journal of Cell Biology, 90, 7-17.
  • Edwards M. K., Wood W. B. 1983. Localization of specific messenger RNAs in C. elegans by cytological hybridization. Developmental Biology, 97, 375-390.
  • Fetterer R. H. 1989. The cuticular proteins from free-living and parasitic stages of Haemonchus contortus. I. Isolation and partial characterization. Comparative Biochemistry and Physiology, 94B, 383-388.
  • Fetterer R. H., Rhodas M. L. 1993. Biochemistry of the nematode cuticle: relevance to parasitic nematodes of livestock. Veterinary Parasitology, 46, 103-111.
  • Fetterer R. H., Hill D. E., Urban J. F., Jr. 1990. The cuticular biology in developmental stages of Ascaris suum. Acta Tropica, 47, 289-295.
  • Hill D. E., Fetterer R. H., Urban J. F., Jr. 1990. Biotin as a probe of the surface Ascaris suum developmental stages. Molecular and Biochemical Parasitology, 41, 45-52.
  • Keith K. A., Duncan M. C., Murray M., Bairden K., Tait A. 1990. Stage-specific cuticular proteins of Ostertagia circumcincta and Ostertagia ostertagi. International Journal for Parasitology, 20, 1037-1045.
  • Laemmli U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680- 685.
  • McBride O. W., Harrington W. F. 1967. Ascaris cuticle collagen: On the disulfide cross-linkages and molecular properties of the subunits. Biochemistry, 6, 1484-1498.
  • Petralanda I., Piessens W. F. 1991. Onchocerca volvulus, O. gutturosa, Brugia malayi and Dirofilaria immitis: A comparative study of the immunochemical properties of the cuticular proteins from filarial parasites. Experimental Parasitology, 72,164— 173.
  • Selkirk M. E., Nielsen L., Kelly C., Partono F., Sayers G., Maizels R. M. 1989. Identification, synthesis and immunogenicity of cuticular collagens from the filarial nematodes Brugia malayi and Brugia pahangi. Molecular and Biochemical Parasitology, 32, 229-246.
  • Shamansky L. M., Pratt D., Boisvenue R. J., Cox G. N. 1989. Cuticle collagen genes of Haemonchus contortus and Caenorhabditis elegans are highly conserved. Molecular and Biochemical Parasitology, 37, 73-86.
  • Towbin H., Stacklin T., Gordon J. 1979. Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America, 76, 4350-4354.
  • Wędrychowicz H., Bairden K., Tait A., Holmes P. H. 1992. Immune response of sheep to surface antigens of infective larvae of Ostertagia circumcincta. Parasite Immunology, 14, 249-266.

Typ dokumentu

Bibliografia

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