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

Palacín Gómez, Aranzazu; Varela, J.; Pozo, V. del; Tordesillas Villuendas, Leticia; Barranco, P.; Fernandez Nieto, M.; Sastre, J.; Díaz Perales, Araceli y 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.

Descripción

Título: Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma
Autor/es:
  • Palacín Gómez, Aranzazu
  • Varela, J.
  • Pozo, V. del
  • Tordesillas Villuendas, Leticia
  • Barranco, P.
  • Fernandez Nieto, M.
  • Sastre, J.
  • Díaz Perales, Araceli
  • Salcedo Duran, Gabriel
Tipo de Documento: Artículo
Título de Revista/Publicación: Clinical and Experimental Allergy
Fecha: Agosto 2009
Volumen: 39
Materias:
Escuela: E.T.S.I. Agrónomos (UPM) [antigua denominación]
Departamento: Biotecnologia [hasta 2014]
Licencias Creative Commons: Reconocimiento - Sin obra derivada - No comercial

Texto completo

[img]
Vista Previa
PDF (Document Portable Format) - Se necesita un visor de ficheros PDF, como GSview, Xpdf o Adobe Acrobat Reader
Descargar (929kB) | Vista Previa

Resumen

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.

Más información

ID de Registro: 5058
Identificador DC: http://oa.upm.es/5058/
Identificador OAI: oai:oa.upm.es:5058
Identificador DOI: 10.1111/j.1365-2222.2009.03280.x
URL Oficial: http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2222.2009.03280.x/abstract
Depositado por: Memoria Investigacion
Depositado el: 25 Nov 2010 12:21
Ultima Modificación: 20 Abr 2016 14:01
  • Open Access
  • Open Access
  • Sherpa-Romeo
    Compruebe si la revista anglosajona en la que ha publicado un artículo permite también su publicación en abierto.
  • Dulcinea
    Compruebe si la revista española en la que ha publicado un artículo permite también su publicación en abierto.
  • Recolecta
  • e-ciencia
  • Observatorio I+D+i UPM
  • OpenCourseWare UPM