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. 1994 Aug 1;13(15):3481–3486. doi: 10.1002/j.1460-2075.1994.tb06654.x

Characterization of a birch pollen allergen, Bet v III, representing a novel class of Ca2+ binding proteins: specific expression in mature pollen and dependence of patients' IgE binding on protein-bound Ca2+.

S Seiberler 1, O Scheiner 1, D Kraft 1, D Lonsdale 1, R Valenta 1
PMCID: PMC395251  PMID: 7520389

Abstract

A cDNA coding for a birch pollen allergen, Bet v III, with significant sequence homology to Ca2+ binding proteins was isolated from an expression cDNA library using serum IgE from a patient who was allergic to pollen. The deduced amino acid sequence of the pollen allergen contained three typical Ca2+ binding sites. Peptides mimicking the Ca2+ binding sites of Bet v III were synthesized and shown to bind 45Ca in blot overlays. The binding of patients' IgE to the recombinant allergen depended on the native protein conformation and protein-bound Ca2+. Depletion of Ca2+ led to a reversible loss of the IgE binding thus representing a conformational IgE epitope adopted by a polypeptide upon Ca2+ binding. By RNA hybridization it was demonstrated that Bet v III is expressed preferentially in mature pollen. Bet v III therefore represents a pollen allergen which because of its unique structural features also belongs to a novel class of Ca2+ binding proteins.

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Selected References

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  1. Baum P., Furlong C., Byers B. Yeast gene required for spindle pole body duplication: homology of its product with Ca2+-binding proteins. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5512–5516. doi: 10.1073/pnas.83.15.5512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Braam J., Davis R. W. Rain-, wind-, and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis. Cell. 1990 Feb 9;60(3):357–364. doi: 10.1016/0092-8674(90)90587-5. [DOI] [PubMed] [Google Scholar]
  3. Breiteneder H., Pettenburger K., Bito A., Valenta R., Kraft D., Rumpold H., Scheiner O., Breitenbach M. The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease resistance response gene. EMBO J. 1989 Jul;8(7):1935–1938. doi: 10.1002/j.1460-2075.1989.tb03597.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Collinge M., Trewavas A. J. The location of calmodulin in the pea plasma membrane. J Biol Chem. 1989 May 25;264(15):8865–8872. [PubMed] [Google Scholar]
  5. Dizhoor A. M., Chen C. K., Olshevskaya E., Sinelnikova V. V., Phillipov P., Hurley J. B. Role of the acylated amino terminus of recoverin in Ca(2+)-dependent membrane interaction. Science. 1993 Feb 5;259(5096):829–832. doi: 10.1126/science.8430337. [DOI] [PubMed] [Google Scholar]
  6. Elsayed S., Bennich H. The primary structure of allergen M from cod. Scand J Immunol. 1975;4(2):203–208. doi: 10.1111/j.1365-3083.1975.tb02618.x. [DOI] [PubMed] [Google Scholar]
  7. Elsayed S., von Bahr-Lindström H., Bennich H. The primary structure of fragment TM2 of allergen M from cod. Scand J Immunol. 1974;3(5):683–686. doi: 10.1111/j.1365-3083.1974.tb01303.x. [DOI] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  9. Fischer R., Koller M., Flura M., Mathews S., Strehler-Page M. A., Krebs J., Penniston J. T., Carafoli E., Strehler E. E. Multiple divergent mRNAs code for a single human calmodulin. J Biol Chem. 1988 Nov 15;263(32):17055–17062. [PubMed] [Google Scholar]
  10. Goldschmidt-Clermont P. J., Kim J. W., Machesky L. M., Rhee S. G., Pollard T. D. Regulation of phospholipase C-gamma 1 by profilin and tyrosine phosphorylation. Science. 1991 Mar 8;251(4998):1231–1233. doi: 10.1126/science.1848725. [DOI] [PubMed] [Google Scholar]
  11. Goldschmidt-Clermont P. J., Machesky L. M., Baldassare J. J., Pollard T. D. The actin-binding protein profilin binds to PIP2 and inhibits its hydrolysis by phospholipase C. Science. 1990 Mar 30;247(4950):1575–1578. doi: 10.1126/science.2157283. [DOI] [PubMed] [Google Scholar]
  12. Heizmann C. W., Hunziker W. Intracellular calcium-binding proteins: more sites than insights. Trends Biochem Sci. 1991 Mar;16(3):98–103. doi: 10.1016/0968-0004(91)90041-s. [DOI] [PubMed] [Google Scholar]
  13. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  14. Hulen D., Baron A., Salisbury J., Clarke M. Production and specificity of monoclonal antibodies against calmodulin from Dictyostelium discoideum. Cell Motil Cytoskeleton. 1991;18(2):113–122. doi: 10.1002/cm.970180206. [DOI] [PubMed] [Google Scholar]
  15. Jena P. K., Reddy A. S., Poovaiah B. W. Molecular cloning and sequencing of a cDNA for plant calmodulin: signal-induced changes in the expression of calmodulin. Proc Natl Acad Sci U S A. 1989 May;86(10):3644–3648. doi: 10.1073/pnas.86.10.3644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kretsinger R. H., Nockolds C. E. Carp muscle calcium-binding protein. II. Structure determination and general description. J Biol Chem. 1973 May 10;248(9):3313–3326. [PubMed] [Google Scholar]
  17. Ling V., Perera I., Zielinski R. E. Primary structures of Arabidopsis calmodulin isoforms deduced from the sequences of cDNA clones. Plant Physiol. 1991 Aug;96(4):1196–1202. doi: 10.1104/pp.96.4.1196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ling V., Zielinski R. E. Cloning of cDNA Sequences Encoding the Calcium-Binding Protein, Calmodulin, from Barley (Hordeum vulgare L.). Plant Physiol. 1989 Jun;90(2):714–719. doi: 10.1104/pp.90.2.714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ling V., Zielinski R. E. Isolation of an Arabidopsis cDNA sequence encoding a 22 kDa calcium-binding protein (CaBP-22) related to calmodulin. Plant Mol Biol. 1993 May;22(2):207–214. doi: 10.1007/BF00014929. [DOI] [PubMed] [Google Scholar]
  20. Lukas T. J., Iverson D. B., Schleicher M., Watterson D. M. Structural characterization of a higher plant calmodulin : spinacia oleracea. Plant Physiol. 1984 Jul;75(3):788–795. doi: 10.1104/pp.75.3.788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maruyama K., Mikawa T., Ebashi S. Detection of calcium binding proteins by 45Ca autoradiography on nitrocellulose membrane after sodium dodecyl sulfate gel electrophoresis. J Biochem. 1984 Feb;95(2):511–519. doi: 10.1093/oxfordjournals.jbchem.a134633. [DOI] [PubMed] [Google Scholar]
  22. Obermeyer G., Weisenseel M. H. Calcium channel blocker and calmodulin antagonists affect the gradient of free calcium ions in lily pollen tubes. Eur J Cell Biol. 1991 Dec;56(2):319–327. [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schaefer W. H., Lukas T. J., Blair I. A., Schultz J. E., Watterson D. M. Amino acid sequence of a novel calmodulin from Paramecium tetraurelia that contains dimethyllysine in the first domain. J Biol Chem. 1987 Jan 25;262(3):1025–1029. [PubMed] [Google Scholar]
  25. Staiger C. J., Goodbody K. C., Hussey P. J., Valenta R., Drøbak B. K., Lloyd C. W. The profilin multigene family of maize: differential expression of three isoforms. Plant J. 1993 Oct;4(4):631–641. doi: 10.1046/j.1365-313x.1993.04040631.x. [DOI] [PubMed] [Google Scholar]
  26. Tilney L. G., Bonder E. M., Coluccio L. M., Mooseker M. S. Actin from Thyone sperm assembles on only one end of an actin filament: a behavior regulated by profilin. J Cell Biol. 1983 Jul;97(1):112–124. doi: 10.1083/jcb.97.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tilney L. G., Inoué S. Acrosomal reaction of Thyone sperm. II. The kinetics and possible mechanism of acrosomal process elongation. J Cell Biol. 1982 Jun;93(3):820–827. doi: 10.1083/jcb.93.3.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Trewavas A., Gilroy S. Signal transduction in plant cells. Trends Genet. 1991 Nov-Dec;7(11-12):356–361. doi: 10.1016/0168-9525(91)90255-o. [DOI] [PubMed] [Google Scholar]
  29. Valenta R., Duchene M., Ebner C., Valent P., Sillaber C., Deviller P., Ferreira F., Tejkl M., Edelmann H., Kraft D. Profilins constitute a novel family of functional plant pan-allergens. J Exp Med. 1992 Feb 1;175(2):377–385. doi: 10.1084/jem.175.2.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Valenta R., Duchêne M., Pettenburger K., Sillaber C., Valent P., Bettelheim P., Breitenbach M., Rumpold H., Kraft D., Scheiner O. Identification of profilin as a novel pollen allergen; IgE autoreactivity in sensitized individuals. Science. 1991 Aug 2;253(5019):557–560. doi: 10.1126/science.1857985. [DOI] [PubMed] [Google Scholar]
  31. Valenta R., Ferreira F., Grote M., Swoboda I., Vrtala S., Duchêne M., Deviller P., Meagher R. B., McKinney E., Heberle-Bors E. Identification of profilin as an actin-binding protein in higher plants. J Biol Chem. 1993 Oct 25;268(30):22777–22781. [PubMed] [Google Scholar]
  32. Valenta R., Vrtala S., Ebner C., Kraft D., Scheiner O. Diagnosis of grass pollen allergy with recombinant timothy grass (Phleum pratense) pollen allergens. Int Arch Allergy Immunol. 1992;97(4):287–294. doi: 10.1159/000236135. [DOI] [PubMed] [Google Scholar]
  33. Vallier P., DeChamp C., Valenta R., Vial O., Deviller P. Purification and characterization of an allergen from celery immunochemically related to an allergen present in several other plant species. Identification as a profilin. Clin Exp Allergy. 1992 Aug;22(8):774–782. doi: 10.1111/j.1365-2222.1992.tb02818.x. [DOI] [PubMed] [Google Scholar]
  34. Vrtala S., Sperr W. R., Reimitzer I., van Ree R., Laffer S., Müller W. D., Valent P., Lechner K., Rumpold H., Kraft D. cDNA cloning of a major allergen from timothy grass (Phleum pratense) pollen; characterization of the recombinant Phl pV allergen. J Immunol. 1993 Nov 1;151(9):4773–4781. [PubMed] [Google Scholar]
  35. van Eerd J. P., Capony J. P., Ferraz C., Pechère J. F. The amino-acid sequence of troponin C from frog skeletal muscle. Eur J Biochem. 1978 Nov 2;91(1):231–242. doi: 10.1111/j.1432-1033.1978.tb20956.x. [DOI] [PubMed] [Google Scholar]

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