Abstract
Pen a 1, the major shrimp allergen from brown shrimp Penaeus aztecus has been identified as the muscle protein tropomyosin. To identify Pen a 1 IgE binding sites, the reactivities of Pen a 1-specific monoclonal antibodies (mAbs) and shrimp-allergic subjects’ IgE to shrimp and homologous mammalian tropomyosins were analyzed. Pen a 1, purified by preparative SDS-PAGE and commercially obtained porcine, bovine and rabbit tropomyosin were cleaved by CNBr or digested by endoproteinases Lys-C, Glu-C, trypsin, Arg-C and chymotrypsin. Reactivities of Pen a 1-specific mAbs and IgE to the resulting peptides were analyzed by dot blot and immunoblotting. The dot blot analysis showed that mAbs and IgE antibodies did not react with any of the mammalian tropomyosins. The immunoblot analysis showed that all Pen a 1 digests bound IgE or mAbs. However, not all peptides in each digest possessed an IgE binding site. IgE binding intensity and frequency varied by subject and peptide digest. IgE and mAb reactivity patterns were similar but no mAb reproduced the IgE binding patterns indicating that subjects’ IgE bound some epitopes that were not recognized by the Pen a 1-specific mAbs. These studies suggest that IgE-binding epitopes are restricted to certain parts of the Pen a 1 molecule, Pen a 1 may have several similar epitopes, and that Pen a 1 epitopes do not appear to be located in the highly homologous parts of the tropomyosin molecule.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
Daul CB, Slattery M, Reese G, Lehrer SB: Identification of the major brown shrimp (Penaeus aztecus) as the muscle protein tropomyosin. Int Arch Allergy Clin Immunol 1994; 105:49–55.
Daul CB, Slattery M, Morgan JE, Lehrer SB: Identification of a common major Crustacea allergen. J Allergy Clin Immunol 1992;89:194.
Halmepuro L, Salvaggio JE, Lehrer SB: Crawfish and lobster allergens: Identification and structural similarities with other Crustacea. Int Arch Allergy Appl Immunol 1987;84:165–72.
Daul CB, Slattery M, Morgan JE, Lehrer SB: Common Crustacea allergens: identification of B cell epi-topes with shrimp-specific monoclonal antibodies; in Kraft D, Sehon A (eds): Molecular Biology and Immunology of Allergens. Boca Raton, CRC Press, 1993, pp 291–294.
Daul CB, Slattery M, Lehrer SB: Shared antigenic/allergenic epitopes between shrimp Pen a I and fruit fly extract. J Allergy Clin Immunol 1993;91:341.
Daul CB, Morgan JE, Waring NP, McCants ML, Hughes J, Lehrer SB: Immunologie evaluation of shrimp-allergic individuals. J Allergy Clin Immunol 1987;80:716–721.
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680–685.
Goding JW: Antibody production by hybridomas. J Immunol Methods 1980;39:285–308.
Kēhler G: The technique of hybridoma production; in Lefkovits I, Pernis B (eds): Immunological Methods, Academic Press, New York, 1981, vol 2, pp 285–298.
Gross E: The cyanogen bromide reaction. Methods Enzymol 1967; 11:238–255.
Schägger H, von Jagow G: Tricine-sodium dodecyl-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 1987; 166:368–379.
Kyhse-Andersen J: Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods 1984; 10:203–209.
Demeulemester C, Peltre G, Laurent M, Pankeleux D, David B: Cyanogen bromide-activated nitrocellulose membranes: Anew tool for immunoprint techniques. Electrophoresis 1987;8:71–73.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Springer Science+Business Media New York
About this chapter
Cite this chapter
Reese, G., Tracey, D., Daul, C.B., Lehrer, S.B. (1996). IGE and Monoclonal Antibody Reactivities to the Major Shrimp Allergen Pen a 1 (Tropomyosin) and Vertebrate Tropomyosins. In: Sehon, A., HayGlass, K.T., Kraft, D. (eds) New Horizons in Allergy Immunotherapy. Advances in Experimental Medicine and Biology, vol 409. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5855-2_31
Download citation
DOI: https://doi.org/10.1007/978-1-4615-5855-2_31
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-7684-2
Online ISBN: 978-1-4615-5855-2
eBook Packages: Springer Book Archive