Complete Amino Acid Sequence Determination of the Major Allergen of Peach (Prunus persica) Pru p1

Abstract

The major protein allergen of peach (Prunus persica), Pru p 1, has recently been identified as a lipid transfer protein (LTP). The complete primary structure of Pru p 1, obtained by direct amino acid sequence and liquid chromatography-mass spectrometry (LC-MS) analyses with the purified protein, is described here. The protein consists of 91 amino acids with a calculated molecular mass of 9178 Da. The amino acid sequence contains eight strictly conserved cysteines, as do all known LTPs, but secondary structure predictions failed to classify the peach 9 kDa protein as an 'all-alpha type', due to the high frequency of amino acids (nine prolines) disrupting alpha helices. Although the sequence similarity with maize LTP is only 63%, out of the 25 amino acids forming the inner surface of the tunnel-like hydrophobic cavity in maize ns-LTP 16 are identical and 7 similar in the peach homolog, supporting the hypothesis of a similar function.

Hypoallergenic Variants of the <i>Parietaria judaica</i> Major Allergen Par j 1: A Member of the Non-Specific Lipid Transfer Protein Plant Family

“…In fact, nsLTPs are a family of highly structurally correlated proteins which can be isolated from a wide range of plants [21]. Some of them have recently been identified as allergens in pollen and plant-derived food [40, 41, 42, 43], so the disruption of the tertiary structure targeting the disulphide bonds should be considered as a strategy widely useful for all the allergenic members of the nsLTP family.…”

Characterization of the T-Cell Epitopes of the Major Peach Allergen Pru p 3

“…Eight peptides, 16–18 amino acids in length with 5 overlapping residues, were synthesized based on the Pru p 3 sequence [8] from the N- to the C-terminal end (1–16, 12–27, 23–38, 34–49, 44–61, 57–72, 68–82, 74–91). Peptides were assembled on a model 433A Applied Biosystem peptide synthesizer, using a stepwise solid-phase fluorenylmethoxycarbonyl method [26,27], onto a preloaded 2-chlorotrityl chloride resin from Novabiochem (Läufelfingen, Switzerland).…”

“…Previously, we and others sequenced [7] and identified [2,8] the major peach allergen Pru p 3 as a molecule with a molecular weight of 9 kDa belonging to the group of lipid transfer proteins (LTPs). In the Mediterranean area, Pru p 3 is recognized by specific serum IgE antibodies in the majority of peach-allergic subjects [2].…”

Characterization of the allergic T-cell response to Pru p 3, the nonspecific lipid transfer protein in peach

“…Pru p 3 has been cloned, sequenced, and produced as a recombinant allergen in different expression systems. [13][14][15][16][17] IgEbinding epitopes of Pru p 3 have been investigated, and its 3-dimensional crystal structure has been described. [18][19][20] However, despite this detailed characterization, information on the T-cell response to nsLTP is limited.…”