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Originally published In Press as doi:10.1074/jbc.M103041200 on May 23, 2001

J. Biol. Chem., Vol. 276, Issue 30, 27959-27966, July 27, 2001
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Ole e 9, a Major Olive Pollen Allergen Is a 1,3-beta -Glucanase
ISOLATION, CHARACTERIZATION, AMINO ACID SEQUENCE, AND TISSUE SPECIFICITY*

Sonia Huecas, Mayte Villalba, and Rosalía RodríguezDagger

From the Departamento de Bioquímica y Biologia Molecular I, Facultad de Química, Universidad Complutense, Avda. Complutense s/n, 28040 Madrid, Spain

Olive pollen allergy is a clinical disorder affecting the human population of Mediterranean areas. A novel major allergen, Ole e 9, has been isolated from olive pollen by gel permeation, hydrophobic affinity, and reverse-phase high performance liquid chromatographies. It is involved in the allergic responses of 65% of patients suffering olive pollinosis. Ole e 9 (molecular mass of 46.4 kDa) displays 1,3-beta -endoglucanase activity (38.9 ± 5.6 mg of glucose released/min × µmol of protein at pH 4.5-6.0 using laminarin as substrate). It is the first 1,3-beta -glucanase, a member of the "pathogenesis-related" protein family, detected in pollen tissue. Seven tryptic peptides of the allergen were sequenced by Edman degradation and used for designing primers to clone the cDNA codifying the protein. Specific cDNA for Ole e 9 was synthesized from total RNA and amplified using the polymerase chain reaction. The allergen sequence showed an open reading frame of 460 amino acids comprising a putative signal peptide of 26 residues. It shows 39, 33, and 32% sequence identity including the catalytic residues when compared with 1,3-beta -glucanases from wheat, willow, and Arabidopsis thaliana, respectively. Northern blot analysis showed that Ole e 9 transcript is specifically expressed in the pollen tissue, and highly conserved counterparts were only detected in taxonomically related pollens.

* This work was supported by Grant PM98/0094 from the Dirección General de Enseñanza Superior e Investigación Científica (M. E. C., Spain).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF249675.

Dagger To whom correspondence should be addressed. Tel.: 34-91-3944260; Fax: 34-91-3944159; E-mail: rrg@bbm1.ucm.es.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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