Identification of polcalcin as a novel allergen of Amaranthus retroflexus pollen

Vakili Moghaddam, M and Fallahpour, M and Mohammadi, M and Rasi Varaee, F and Mokhtarian, K and Khoshmirsafa, M and Jafari, R and Shirzad, N and Falak, R (2019) Identification of polcalcin as a novel allergen of Amaranthus retroflexus pollen. Allergologia et Immunopathologia.

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Official URL: https://www.sciencedirect.com/science/article/pii/...

Abstract

Introduction: Amaranthus retroflexus (Redroot Pigweed) is one of the main sources of allergenic pollens in temperate areas. Polcalcin is a well-known panallergen involved in cross-reactivity between different plants. The aim of this study was the molecular cloning and expression of polcalcin, as well as evaluating its IgE-reactivity with A. retroflexus sensitive patients’ sera. Methods: Allergenic extract was prepared from A. retroflexus pollen and the IgE-reactivity profile was determined by ELISA and immunoblotting using sera from twenty A. retroflexus sensitive patients. Polcalcin-coding sequence was amplified by conventional PCR method and the product was inserted into pET-21b(+) vector. The recombinant protein was expressed in E. coli BL21 and purified by metal affinity chromatography. The IgE-binding capability of the recombinant protein was analyzed by ELISA and immunoblotting assays, and compared with crude extract. Results: Of 20 skin prick test positive patients, 17 patients were positive in IgE-specific ELISA. Western blotting confirmed that approximately 53% of ELISA positive patients reacted with 10 kDa protein in crude extract. The A. retroflexus polcalcin gene, encoding to 80 amino acid residues was cloned and expressed as a soluble protein and designated as Ama r 3. The recombinant polcalcin showed rather identical IgE-reactivity in ELISA and western blotting with 10 kDa protein in crude extract. These results were confirmed by inhibition methods, too. Conclusion: The recombinant form of A. retroflexus polcalcin (Ama r 3) could be easily produced in E. coli in a soluble form and shows rather similar IgE-reactivity with its natural counterpart

Item Type: Article
Uncontrolled Keywords: Allergen characterization, Amaranthus retroflexus, Cloning, Polcalcin
Subjects: WB Practice of Medicine
QV pharmacology
Divisions: Faculty of Medicine
Depositing User: Unnamed user with email zamani.m@skums.ac.ir
Date Deposited: 17 Feb 2019 05:58
Last Modified: 10 Apr 2019 07:56
URI: http://eprints.skums.ac.ir/id/eprint/7550

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