Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma

Palacín Gómez, Aranzazu and Varela, J. and Pozo, V. del and Tordesillas Villuendas, Leticia and Barranco, P. and Fernandez Nieto, M. and Sastre, J. and Díaz Perales, Araceli and Salcedo Duran, Gabriel (2009). Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma. "Clinical and Experimental Allergy", v. 39 (n. 8); pp. 1267-1276. ISSN 0954-7894. https://doi.org/10.1111/j.1365-2222.2009.03280.x.

Description

Title: Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma
Author/s:
  • Palacín Gómez, Aranzazu
  • Varela, J.
  • Pozo, V. del
  • Tordesillas Villuendas, Leticia
  • Barranco, P.
  • Fernandez Nieto, M.
  • Sastre, J.
  • Díaz Perales, Araceli
  • Salcedo Duran, Gabriel
Item Type: Article
Título de Revista/Publicación: Clinical and Experimental Allergy
Date: August 2009
Volume: 39
Subjects:
Faculty: E.T.S.I. Agrónomos (UPM) [antigua denominación]
Department: Biotecnologia [hasta 2014]
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

BACKGROUND Baker's asthma is an important occupational allergic disease. Wheat lipid transfer protein (LTP) Tri a 14 is a major allergen associated with wheat allergy. No panel of wheat recombinant allergens for component-resolved diagnosis of baker's asthma is currently available. OBJECTIVE To evaluate the potential role of recombinant Tri a 14 as a novel tool for the diagnosis of baker's asthma, and to test the heat and proteolytic resistance of the wheat LTP allergen. METHODS A cDNA encoding Tri a 14 was isolated and sequenced, the recombinant allergen produced in Pichia pastoris and purified by chromatographic methods. Physicochemical and immunological comparison of the natural and recombinant forms of Tri a 14 was carried out by N-terminal amino acid sequencing, matrix-assisted laser desorption/ionization mass spectrometry, circular dichroism (CD) analysis, IgE immunodetection, and specific IgE determination and ELISA-inhibition assays using a pool or individual sera from 26 patients with baker's asthma. Thermal denaturation and simulated gastrointestinal digestion of both Tri a 14 forms were checked by spectroscopic and electrophoretic methods, respectively, and biological activity by basophil activation test (BAT). RESULTS Natural and recombinant Tri a 14 were similarly folded, as indicated by their nearly identical CD spectra and heat denaturation profiles. A high interclass correlation coefficient (0.882) was found between specific IgE levels to both Tri a 14 proteins in individual sera from baker's asthma patients, but a slightly lower IgE-binding potency of rTri a 14 was detected by ELISA-inhibition assays. Natural and recombinant Tri a 14 elicited positive BAT in two and one out of three patients, respectively. Heat denaturation profiles and simulated gastrointestinal digestion assays indicated that Tri a 14 displayed a high heat and digestive proteolytic resistance, comparable to those of peach Pru p 3, the model food allergen of the LTP family. CONCLUSIONS Recombinant Tri a 14 is a potential tool for baker's asthma diagnosis, based on its physicochemical and immunological similarity with its natural counterpart. Wheat Tri a 14 shows a high thermal stability and resistance to gastrointestinal digestion.

More information

Item ID: 5058
DC Identifier: http://oa.upm.es/5058/
OAI Identifier: oai:oa.upm.es:5058
DOI: 10.1111/j.1365-2222.2009.03280.x
Official URL: http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2222.2009.03280.x/abstract
Deposited by: Memoria Investigacion
Deposited on: 25 Nov 2010 12:21
Last Modified: 20 Apr 2016 14:01
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