Identification of a polygalacturonase as a major allergen (Pla a 2) from Platanus acerifolia pollen

被引:78
作者
Ibarrola, I [1 ]
Arilla, MC [1 ]
Martínez, A [1 ]
Asturias, JA [1 ]
机构
[1] Bial Aristegui, Dept Res & Dev, Bilbao 48008, Spain
关键词
London planetree; major allergen; Platanus acerifolia; polygalacturonase; three-dimensional structure; molecular modeling;
D O I
10.1016/j.jaci.2004.02.031
中图分类号
R392 [医学免疫学];
学科分类号
100102 ;
摘要
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. Conclusion: Pla a 2, a major allergen of P acerifolia pollen with PG activity has been purified, characterized, and cloned.
引用
收藏
页码:1185 / 1191
页数:7
相关论文
共 27 条
[1]  
Aalberse RC, 2000, J ALLERGY CLIN IMMUN, V106, P228, DOI 10.1067/mai.2000.108434
[2]   The major Platanus acerifolia pollen allergen Pla a 1 has sequence homology to invertase inhibitors [J].
Asturias, JA ;
Ibarrola, I ;
Eraso, E ;
Arilla, MC ;
Martínez, A .
CLINICAL AND EXPERIMENTAL ALLERGY, 2003, 33 (07) :978-985
[3]   Purification and characterization of Pla a 1, a major allergen from Platanus acerifolia pollen [J].
Asturias, JA ;
Ibarrola, I ;
Bartolomé, B ;
Ojeda, I ;
Malet, A ;
Martínez, A .
ALLERGY, 2002, 57 (03) :221-227
[4]   Worldwide variation in prevalence of symptoms of asthma, allergic rhinoconjunctivitis, and atopic eczema:: ISAAC [J].
Beasley, R ;
Keil, U ;
von Mutius, E ;
Pearce, N ;
Aït-Khaled, N ;
Anabwani, G ;
Anderson, HR ;
Asher, MI ;
Björkstéin, B ;
Burr, ML ;
Clayton, TO ;
Crane, J ;
Ellwood, P ;
Lai, CKW ;
Mallol, J ;
Martinez, FD ;
Mitchell, EA ;
Montefort, S ;
Robertson, CF ;
Shah, JR ;
Sibbald, B ;
Stewart, AW ;
Strachan, DP ;
Weiland, SK ;
Williams, HC .
LANCET, 1998, 351 (9111) :1225-1232
[5]   Purification and characterization of an 18-kd allergen of birch (Betula verrucosa) pollen:: Identification as a cyclophilin [J].
Cadot, P ;
Díaz, JF ;
Proost, P ;
Van Damme, J ;
Engelborghs, Y ;
Stevens, EAM ;
Ceuppens, JL .
JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 2000, 105 (02) :286-291
[6]   NEW AND SIMPLE RADIOIMMUNOASSAY METHOD FOR DETERMINATION OF IGE [J].
CESKA, M ;
LUNDKVIST, U .
IMMUNOCHEMISTRY, 1972, 9 (10) :1021-+
[7]   Polygalacturonase (pectinase), a new oilseed rape allergen [J].
Chardin, H ;
Mayer, C ;
Sénéchal, H ;
Poncet, P ;
Clément, G ;
Wal, JM ;
Desvaux, FX ;
Peltre, G .
ALLERGY, 2003, 58 (05) :407-411
[8]  
Charpin J., 1974, ATLAS EUROPEAN ALLER
[9]   THE PARALLEL BETA-HELIX OF PECTATE LYASE-C - SOMETHING TO SNEEZE AT [J].
COHEN, FE .
SCIENCE, 1993, 260 (5113) :1444-1445
[10]   DETERMINATION OF REDUCING SUGAR WITH IMPROVED PRECISION [J].
DYGERT, S ;
LI, LH ;
FLORIDA, D ;
THOMA, JA .
ANALYTICAL BIOCHEMISTRY, 1965, 13 (03) :367-&