cDNA cloning, expression and primary structure of Par j I, a major allergen of Parietaria judaica pollen

Abstract

A 659 bp cDNA clone** coding for an allergen of Pj pollen has been isolated from a lambda gt 11 library, and its DNA sequence determined. The cDNA insert showed an open reading frame of 429 bp coding for an allergenic protein of 14,866 Da and a deduced amino acid sequence containing 143 residues. The expressed recombinant protein represented the major allergen Par j I since it reacted with 95% of the sera from Pj-allergic patients (n = 22) and with two Par j I-specific monoclonal antibodies. No similarity with other known DNA and protein sequences has been detected.

An approach to the knowledge of pollen and allergen diversity through lipid transfer protein localisation in taxonomically distant pollen grains

“…It is interesting to point out that this labelling was less intense than that obtained with Pla a 1 and Pla a 2, the major allergens of P. acerifolia pollen grains (Suárez-Cervera et al, 2005). Regarding P. judaica pollen grains, their major allergens, Par j 1 and Par j 2, represent another type of nsLTP member (Ayuso et al, 1995;Costa et al, 1994;Asturias et al, 2003). Although the sequential positions for IgE epitopes of Par j 1 and Par j 2 and Pru p 3 show similarities of 26-29%, both allergens have strong sequence divergence in most of their regions (Salcedo et al, 2004(Salcedo et al, , 2007.…”

“…Recently published reviews have provided a thorough background on nsLTPs in foods and pollen (Mills et al, 2004;Salcedo et al, 2004Salcedo et al, , 2007Breiteneder & Mills, 2005). Major allergens of Parietaria judaica L. pollen grains, Par j 1 and Par 2, belong to the nsLTP family (Costa et al, 1994;Asturias et al, 2003; (1). Fumariaceae, Fumaria capreolata (18.5 mm); (2).…”

Fitotoxicidad De Aceites Esenciales Y Extractos Acuosos De Plantas Mediterráneas Para El Control De Arvenses

“…judaica es una de las fuentes más importante de polen alérgeno en la zona Mediterránea (Stumvoll et al, 2003;Fotiu et al, 2011). La mayoría de trabajos existentes sobre esta especie han sido llevados a cabo para caracterizar los alérge-nos presentes en su polen (Ford et al, 1986;Costa et al, 1994;Duro et al, 1996;Stumvoll et al, 2003), además de estudiar distintos factores que condicionan la concentración del mismo en el ambiente (Fornaciari et al, 1992;Galán et al, 2000;Fotiu et al, 2011).…”

Amino Acid Substitution in Par j 2 Recombinant Allergen and Its Effect on IgE Binding Capacity

“…These proteins possess a preserved secondary structure that forms a motif α-α-α-α-β and contain 8 cysteines that form 4 disulphuric bridges that stabilize the three-dimensional structure. [3][4][5][6][7][8].…”

“…In our previous studies, these regions have been highlighted as important IgE binding epitopes [4,6] and we have seen that Par j 2 regions 1-30, 30-55, and 77-102 were capable of binding specific IgE from sera of allergic patients to PJ (Figure 1), [4,9]. To identify potential amino acid residues directly involved in IgE binding, site-directed mutagenesis was performed on the region 38-55 [3][4][5][6]. The mutants prepared by site-directed mutagenesis were named K41 (2399.66 Da), T42 (2426.73 Da), T43 (2426.73 Da), C50 (2424.69 Da), and C52 (2424.69 Da), where alanine was the amino acid which was replaced.…”