The crystal structure of recombinant proDer p 1, a major house dust mite proteolytic allergen

TY - JOUR

T1 - The crystal structure of recombinant proDer p 1, a major house dust mite proteolytic allergen

AU - Meno, Kåre

AU - Thorsted, Peter B

AU - Ipsen, Henrik

AU - Kristensen, Ole

AU - Larsen, Jørgen

AU - Spangfort, Michael D

AU - Gajhede, Michael

AU - Lund, Kaare

PY - 2005

Y1 - 2005

N2 - Allergy to house dust mite is among the most prevalent allergic diseases worldwide. Most house dust mite allergic patients react to Der p 1 from Dermatophagoides pteronyssinus, which is a cysteine protease. To avoid heterogeneity in the sample used for crystallization, a modified recombinant molecule was produced. The sequence of the proDer p 1 allergen was modified to reduce glycosylation and to abolish enzymatic activity. The resulting rproDer p 1 preparation was homogenous and stable and yielded crystals diffracting to a resolution of 1.61 A. The active site is located in a large cleft on the surface of the molecule. The 80-aa pro-peptide adopts a unique fold that interacts with the active site cleft and a substantial adjacent area on the mature region, excluding access to the cleft and the active site. Studies performed using crossed-line immunoelectrophoresis and IgE inhibition experiments indicated that several epitopes are covered by the pro-peptide and that the epitopes on the recombinant mature molecule are indistinguishable from those on the natural one. The structure confirms previous results suggesting a preference for aliphatic residues in the important P2 position in substrates. Sequence variations in related species are concentrated on the surface, which explains the existence of cross-reacting and species-specific antibodies. This study describes the first crystal structure of one of the clinically most important house dust mite allergens, the cysteine protease Der p 1.

AB - Allergy to house dust mite is among the most prevalent allergic diseases worldwide. Most house dust mite allergic patients react to Der p 1 from Dermatophagoides pteronyssinus, which is a cysteine protease. To avoid heterogeneity in the sample used for crystallization, a modified recombinant molecule was produced. The sequence of the proDer p 1 allergen was modified to reduce glycosylation and to abolish enzymatic activity. The resulting rproDer p 1 preparation was homogenous and stable and yielded crystals diffracting to a resolution of 1.61 A. The active site is located in a large cleft on the surface of the molecule. The 80-aa pro-peptide adopts a unique fold that interacts with the active site cleft and a substantial adjacent area on the mature region, excluding access to the cleft and the active site. Studies performed using crossed-line immunoelectrophoresis and IgE inhibition experiments indicated that several epitopes are covered by the pro-peptide and that the epitopes on the recombinant mature molecule are indistinguishable from those on the natural one. The structure confirms previous results suggesting a preference for aliphatic residues in the important P2 position in substrates. Sequence variations in related species are concentrated on the surface, which explains the existence of cross-reacting and species-specific antibodies. This study describes the first crystal structure of one of the clinically most important house dust mite allergens, the cysteine protease Der p 1.

KW - Allergens

KW - Animals

KW - Arthropod Proteins

KW - Binding Sites

KW - Cross Reactions

KW - Crystallography, X-Ray

KW - Epitopes

KW - Genetic Engineering

KW - Models, Molecular

KW - Molecular Structure

KW - Protein Conformation

KW - Protein Precursors

KW - Pyroglyphidae

KW - Recombinant Proteins

KW - Species Specificity

KW - Substrate Specificity

M3 - Journal article

C2 - 16148130

SN - 0022-1767

VL - 175

SP - 3835

EP - 3845

JO - Journal of Immunology

JF - Journal of Immunology

IS - 6

ER -