Molecular basis of IgE-recognition of Lol p 5, a major allergen of rye-grass pollen.
Grass pollen, especially of rye-grass (Lolium perenne). represents an important cause of type I allergy. Identification of IgE-binding (allergenic) epitopes of major grass pollen allergens is essential for understanding the molecular basis of interaction between allergens and human IgE antibodies and therefore facilitates the devising of safer and more effective diagnostic and immunotherapy reagents. The aim of this study was to identify the allergenic epitopes of Lol p 5, a major allergen of rye-grass pollen, immunodissect these epitopes further so that the amino acid residues critical for antibody binding can be determined and investigate the conservation and nature of these epitopes within the context of the natural grass pollen allergens. Peptides, 12-13 amino acid residues long and overlapping each other by 4 amino acid residues, based on the entire deduced amino acid sequence of the coding region of Lol p 5, were synthesised and assayed for IgE-binding. Two strong IgE-binding epitopes (Lol p 5 (49-60) and (265-276), referred to as peptides 7 and 34, respectively) were identified. These epitopes were further resolved by truncated peptides and amino acid replacement studies and the amino acid residues critical for IgE-binding determined (Lol p 5 (49-60) residue Lys57 and (265-276) residue Lys275). Sequences of these epitopes were conserved in related allergens and may form the conserved allergenic domains responsible for the cross-reactivity observed between pollen allergens of taxonomically related grasses. Furthermore, due to its strong IgE-reactivity, synthetic peptide Lol p 5 (265-276) was used to affinity-purify specific IgE antibodies which recognised proteins of other clinically important grass pollens. further indicating presence of allergenic cross-reactivity at the level of allergenic epitope. Moreover, Lol p 5 (265 276) demonstrated a strong capacity to inhibit IgE-binding to natural rye-grass pollen proteins highlighting the antibody accessibility to these sequences within the context of the natural allergens. Strong IgE-binding epitopes of Lol p 5 have been identified down to single critical amino acid residues and are shown to occur as linear or continuous domains in the natural conformation of natural Lol p 5 and other group 5 grass pollen allergens. The fact that such an allergenic synthetic epitope has the capacity to strongly inhibit IgE-binding to natural allergens highlight its potential for use as a candidate in future therapeutics to treat pollen-associated allergies.
Suphioglu
C
C
School of Botany, University of Melbourne, Parkville, Victoria, Australia. cenk.suphioglu@med.monash.edu.au
Blaher
B
B
Rolland
J M
JM
McCluskey
J
J
Schäppi
G
G
Kenrick
J
J
Singh
M B
MB
Knox
R B
RB
D000485:Allergens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001666:Binding Sites, Antibody; D017124:Conserved Sequence; D000939:Epitopes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism*; D008129:Lolium / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D011058:Pollen / Q000737:chemistry / Q000276:immunology*; D017386:Sequence Homology, Amino Acid
D000485:Allergens; D052179:Antigens, Plant; D000939:Epitopes; C084941:Lol p V protein, Lolium perenne; D009842:Oligopeptides; D010940:Plant Proteins; D007073:Immunoglobulin E