Identification of a polygalacturonase as a major allergen (Pla a 2) from Platanus acerifolia pollen.
BACKGROUND
Planetree pollen allergy is a clinical disorder affecting human populations in cities of the United States and Western Europe, but little is known about its relevant allergens.


OBJECTIVE
We sought to purify, characterize, and clone the 43-kd allergen from Platanus acerifolia.


METHODS
P acerifolia pollen extract was fractionated by using ion-exchange and gel-permeation chromatography. Analyses were carried out by using ELISA, SDS-PAGE, isoelectrofocusing, and immunoblotting. Partial amino acid sequence was obtained by means of Edman sequencing of cyanogen bromide-digested peptides. Specific cDNA was cloned by using reverse transcription, followed by PCR, with amino acid sequences from peptides of the allergen.


RESULTS
The allergen isolated from P acerifolia pollen, Pla a 2, is a glycoprotein with an observed molecular mass of 43 kd and an isoelectric point value of 9.3. It is involved in the allergic responses of 84% of patients with planetree-induced pollinosis and represented 52% of the total IgE-binding capacity of the P acerifolia extract. Pla a 2 displays polygalacturonase (PG) activity, being the first PG with functional enzyme activity from an angiosperm plant pollen described as an allergen. The cDNA allergen sequence codified for a 372-residue protein with 56% and 42% sequence identity to PGs from pollen and fruits, respectively. Western blot analysis showed that Pla a 2 is present in pollen and stems and has IgG cross-reactivity with a PG from tomato and pectate lyases from Cupressaceae pollen.


CONCLUSIONS
Pla a 2, a major allergen of P acerifolia pollen with PG activity has been purified, characterized, and cloned.
Ibarrola
Ignacio
I
Research and Development Department, Bial-Arstegui, Alameda Urquijo 27, 48008-Bilbao, Spain.
Arilla
M Carmen
MC
Martínez
Alberto
A
Asturias
Juan A
JA
D000328:Adult; D000485:Allergens / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D011058:Pollen / Q000276:immunology*; D011096:Polygalacturonase / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits; D014197:Trees / Q000276:immunology*
D000485:Allergens; D011096:Polygalacturonase