Identification of a 53 kDa protein, as a new high molecular weight allergen from Fraxinusexcelsior (Ash) pollen

Summary

Background

Fraxinus excelsior (Ash) is a common tree and is important cause of winter–spring pollinosis in many temperate regions in the world. In this study, a high molecular weight allergen from ash pollen was identified.

Methods

In all, 20 individuals allergic to ash participated in the study. Characterization and immunoreactivity of ash pollen proteins was performed using sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE), two-dimensional (2D) gel electrophoresis and immunoblotting.

Results

Immunoglobulin E (IgE)-binding proteins with apparent molecular mass ranging from 9 to 110 kDa were detected in ash pollen extract. Serum IgE of 7 (35%) patients reacted with a 53-kDa protein band. Analysis of 2D immunoblots showed several IgE-binding proteins. Moreover, mass spectrometry analysis indicated that the 53-kDa allergen was homologous to calreticulin.

Discussion

We defined a novel allergen from F. excelsior pollen with a molecular weight of about 53 kDa. This allergen could be considered as an important high molecular weight allergenic protein for further studies on cross-reactivity and development of diagnostic and therapeutic approaches.

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Abbreviations

2‑DE:

Two-dimensional electrophoresis

BSA:

Bovine serum albumin

CBB:

Comassie brilliant blue

CRT:

Calreticulin

ER:

Endoplasmic reticulum

HMW:

High molecular weight

IAARI:

Immunology, Asthma and Allergy Research Institute

IEF:

Isoelectric focusing

IPG:

Immobilized pH gradient

IPI:

International protein index

LiquidDAB+:

High-sensitivity liquid diaminobenzidine

MALDI:

Matrix assisted laserdesorption/ionization

MW:

Molecular weight

PVDF:

Polyvinylidene fluoride

SDS-PAGE:

SDS-polyacrylamide gel electrophoresis

TBS:

Tris-buffered saline

TOF:

Time of flight

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Acknowledgements

This study was supported by Immunology, Asthma and Allergy Research Institute (IAARI), Children’s Medical Center, Tehran University of Medical Sciences (grant # 93-03-40-27007).

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Correspondence to Gholam Ali Kardar PhD.

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Conflict of interest

M. Sharif Shoushtari, A. Majd, M.-A. Assarehzadegan, S. Fanuel, M. Moin, T. Nejadsattari, R. S. Shoormasti,M. Badalzadeh, S. Tajik, M.R. Fazlollahi, B. Tayebi, Z. Pourpak and G.A. Kardar declare that they have no competing interests.

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Sharif Shoushtari, M., Majd, A., Assarehzadegan, M. et al. Identification of a 53 kDa protein, as a new high molecular weight allergen from Fraxinusexcelsior (Ash) pollen. Allergo J Int (2020). https://doi.org/10.1007/s40629-020-00129-3

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Keywords

  • Ash pollen
  • Allergen
  • 2D electrophoresis
  • IgE immunoblotting
  • Calreticulin