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Isolation and partial characterisation of the serum immunoglobulin of the flounder, Platichthys flesus

Published online by Cambridge University Press:  11 May 2009

P.J. Glynn  and
A.L. Pulsford
P.J. Glynn
Affiliation:
Department of Biological Sciences, Polytechnic South West, Plymouth, PL4 8AA
A.L. Pulsford
Affiliation:
Plymouth Marine Laboratory, Citadel Hill, Plymouth, PL1 2PB
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Abstract

Whole flounder serum has been fractionated by gel filtration on Ultrogel AcA22. Pooled material from the first major peak of the elution profile was further fractionated by FPLC ion-exchange chromatography and this yielded one major and three minor peaks. Under both reducing and non-reducing conditions SDS electrophoresis showed that the major peak and two of the minor ones were immunoglobulin (Ig). All three of these Ig populations were tetrameric, with estimated molecular weights of 710 kD. However, approximately 90% of the major Ig species consisted of tetramers which were covalently linked by disulphide bonds, whereas the remaining 10% was composed of dimeric molecules held together by non-covalent interactions. The heavy chains of all three Ig populations had apparent molecular weights of approximately 72 kD but the light chain composition showed considerable heterogeneity. In the major population, five polypep-tides were detected in the light chain region of the gel with apparent molecular weights covering the range 22–28 kD. However, there was no difference in the light chain composition of covalently and non-covalently linked tetramers. The two minor populations differed both from each other and from the major species in respect of their light chain compositions. No evidence was found for a monomeric serum Ig, and alkaline urea electrophoresis failed to demonstrate a J chain.

Type
Research Article
Information
Journal of the Marine Biological Association of the United Kingdom , Volume 70 , Issue 2 , May 1990 , pp. 429 - 440
Copyright
Copyright © Marine Biological Association of the United Kingdom 1990

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References

Acton, R.T., Weinheimer, P.F., Hall, S.J., Niedermeier, W., Shelton, E. & Bennett, J.C. 1971. Tetrameric immune macroglobulins in three orders of bony fishes. Proceedings of the National Academy of Sciences of the United States of America, 68, 107–111.CrossRefGoogle ScholarPubMed
Bradford, M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein. Analytical Biochemistry, 72, 248.CrossRefGoogle ScholarPubMed
Clem, L.W. & McLean, W.E. 1975. Phylogeny of immunoglobulin structure and function. VII. Monomeric and tetrameric immunoglobulins of the margate, a marine teleost fish. Immunology, 29, 791–799.Google ScholarPubMed
Dorson, M., 1981. Role and characterisation of fish antibody. Developments in Biological Standardization, 49, 307–319.Google Scholar
Frommel, D., Litman, G.W., Finstadd, J. & Good, R.A., 1971. The evolution of the immune response. Journal of Immunology, 106, 1234–1243.CrossRefGoogle ScholarPubMed
Havarstein, S.L., Aasjord, P.M., Ness, S. & Endresen, C., 1988. Purification and partial characterisation of an IgM-like serum immunoglobulin from Atlantic salmon (Salmo salar). Developmental and Comparative Immunology, 12, 773–785.CrossRefGoogle ScholarPubMed
Johnston, W.H. Jr, Acton, R.T., Weinheimer, P.F., Niedermeier, W., Evans, E.E., Shelton, E. & Bennett, J.C., 1971. Isolation and physico-chemical characterisation of the ‘IgM-like’ immunoglobulin from the stingray Dasyatis americana. Journal of Immunology, 107, 782–793.CrossRefGoogle ScholarPubMed
Johnstone, A. & Thorpe, R., 1987. lmmunochemistry in Practice, 2nd ed. Oxford: Blackwell Scientific Publications.Google Scholar
Kobayashie, K., Hara, A., Takano, K. & Hirai, H., 1982. Studies on subunit components of immunoglobulin M from a bony fish, the chum salmon (Oncorhynchus keta). Molecular Immunology, 19,95–103.Google Scholar
Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London, 227, 680–685.CrossRefGoogle ScholarPubMed
Litman, G.W., 1976. Physical properties of immunoglobulins of lower species: a comparison with immunoglobulins of mammals. In Comparative Immunology (ed. J.J., Marchalonis), pp. 239–275. Oxford: Blackwell Scientific Publications.Google Scholar
Lobb, C.J. & Clem, L.W., 1981 a. The metabolic relationships of the immunoglobulins in fish serum, cutaneous mucus and bile. Journal of Immunology, 127, 1525–1529.CrossRefGoogle ScholarPubMed
Lobb, C.J. & Clem, L.W., 1981 b. Phylogeny of immunoglobulin structure and function. X. Humoral immunoglobulins of the sheepshead Archosargus probatocephalus. Developmental and Comparative Immunology, 5, 271–282.CrossRefGoogle ScholarPubMed
Lobb, C.J. & Clem, L.W., 1981 c. Phylogeny of immunoglobulin structure and function. XI. Secretory immunoglobulins in the cutaneous mucus of the sheepshead Archosargus probatocephalus. Developmental and Comparative Immunology, 5, 587–596.CrossRefGoogle ScholarPubMed
Lobb, C.J. & Clem, L.W., 1983. Distinctive subpopulations of catfish serum antibody and immunoglobulin. Molecular Immunology, 120, 811–818.CrossRefGoogle Scholar
Lobb, C.J., Olson, M.O. & Clem, L.W., 1984. Immunoglobulin light chain classes in a teleost fish. Journal of Immunology, 132, 1917–1923.CrossRefGoogle Scholar
McCumber, L.J. & Clem, L.W., 1976. A comparative study of J chain structure and stoichiometry in human and nurse shark IgM. lmmunochemistry, 13, 479–484.CrossRefGoogle ScholarPubMed
Marchalonis, J.J. 1977. Emergence of IgM immunoglobulins. In Immunity and Evolution. London: Edward Arnold.Google Scholar
Morrissey, J., 1981. Silver stain for proteins in polyacrylamide gels. A modified procedure with enhanced uniform sensitivity. Analytical Biochemistry, 117, 307–310.CrossRefGoogle ScholarPubMed
Sanchez, C., Dominguez, J. & Coli, J., 1989. Immunoglobulin heterogeneity in the rainbow trout, Salmo gairdneri Richardson. Journal of Fish Diseases, 12, 459–465.CrossRefGoogle Scholar
Tomonaga, S., Kobayashi, K., Hagiwara, K., Sasaki, K. & Sezaki, K., 1985. Studies on immunoglobulin and immunoglobulin-forming cells in Heterodontus japonicus, a cartilaginous fish. Developmental and Comparative Immunology, 9,617–626.CrossRefGoogle ScholarPubMed
Weinheimer, P.F., Mestecky, J. & Acton, R.T., 1971. Species distribution of J chain. Journal of Immunology, 107, 1211–1212.CrossRefGoogle ScholarPubMed