Molecular cloning of a house dust mite allergen with common antibody binding specificities with multiple components in mite extracts

Hd, Shen; Chua, Kevin Yiqiang; Lin, Kun; Kh, Hsieh; Wr, Thomas

doi:10.1111/j.1365-2222.1993.tb00278.x

Cited by 57 publications

(74 citation statements)

References 22 publications

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“…The interpretation of the studies is however difficult because of variation from report to report, the poor dose-response characteristics of blotting and the lability of allergens. Recent studies have also shown that the group 3, 7 and 14 allergens are found with multiple bands which vary from extract to extract [11, 12, 13]. The use of recombinant techniques has provided a method of examining the allergens without relying on their abundance or stability in extracts.…”

Section: House Dust Mite Extractsmentioning

confidence: 99%

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Characterization and Immunobiology of House Dust Mite Allergens

Thomas¹,

Smith²,

Hales³

et al. 2002

Int Arch Allergy Immunol

303

254

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The examination of house dust mite extracts has indicated that over 30 different proteins can induce IgE antibody in patients allergic to the house dust mite. There are however dominant specificities especially the group 1 and 2 allergens which can account for much of the allergenicity of extracts. Of the 19 denominated allergens, the major IgE binding has been reported for the group 1, 2, 3, 9, 11, 14 and 15 allergens. The high-molecular-weight group 11, 14 and 15 allergens have only recently been described and although high IgE binding has been anticipated from immunoblotting, there is a need for considerable corroboration. Similarly, the study of the group 3 and 9 serine protease allergens has been incomplete. The group 4, 5, 7 and 8 allergens have shown intermediate IgE binding and the group 10 tropomyosins are of interest because of their potential cross-reactivity with allergen from disparate species. Although the progress with the production of recombinant group 1 allergens has been recent, many of the allergens can be produced as high IgE-binding polypeptides. The tertiary structure of the group 2 allergens has been determined from recombinant proteins and they are an excellent model for the investigation of modified allergens. An unexpected property of the group 1, 2 and 3 allergens has been the high degree of polymorphism found by cDNA analysis. It has however been possible to identify sequences to represent the variation in the natural allergens. The group 7 and 14 allergens show secondary modifications which vary in different extracts creating batch variation. While some estimate of the importance of allergens can be obtained from IgE binding, few analyses of T-cell responses have been made and these regulate both the development of, and the protection from sensitization.

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Section: House Dust Mite Extractsmentioning

confidence: 99%

“…The cDNA encoded 215 residues with no cysteines, a single N-glycosylation site and a predicted 17-residue leader peptide [12]. The mature 198-residue polypeptide had a MW of 22,000.…”

Section: Group 7 Allergensmentioning

confidence: 99%

Characterization and Immunobiology of House Dust Mite Allergens

Thomas¹,

Smith²,

Hales³

et al. 2002

Int Arch Allergy Immunol

303

254

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“…Considerable information is available on many of the major mite allergens, particularly those of the genus Dermatopha goides, either through physicochemical studies of the native proteins [7][8][9][10], or the analysis of the cDNA coding for these, and the evaluation of the expression products of this cDNA [11,12], A particularly important allergen of Dermatophagoidespteronyssinus, designated Dcr p 2, has been ex tensively studied [13,14], and has been cloned and ex pressed in Escherichia coli as a fusion protein with glutathione-S-transferase (GST) [15,16]. We have shown that the recombinant material is closely comparable to the native protein in its capacity to bind specific IgE and to elicit im mediate hypersensitivity reactions in the skin o f house-dust mite-allergic patients [17], Two less well characterized mite allergens, Der p 5 [ 18,19] and Der p 7 [20], have also been cloned, and the recombinant fusion proteins shown to bind IgE and produce positive skin tests.…”

Section: Introductionmentioning

confidence: 99%

Biological Activity of Recombinant Der p 2, Der p 5 and Der p 7 Allergens of the House-Dust Mite <i>Dermatophagoides pteronyssinus</i>

Lynch

Thomas

García

et al. 1997

Int Arch Allergy Immunol

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Der p 2, Der p 5 and Der p 7 are three allergens of the house-dust mite Dermatophagoides pteronyssinus that have been cloned and expressed in Escherichia coli as fusion proteins with glutathione-S-transferase (GST). We showed that these recombinant allergens produced immediate hypersensitivity skin-test reactions in 70, 60 and 52% respectively of a group of mite-sensitive allergic patients who were strongly positive to whole mite extract (WME). Comparable positiv-ities were found for serum levels of specific IgE antibody against these allergens, as measured by the radioallergosorbant test (RAST). Overall, for the group of allergic patients that we evaluated, the serum IgE antibody concentrations against Der p 2, 5 and 7 were calculated to represent about one third, one quarter and one fifth respectively of the levels measured against the WME. However, for some patients the activity determined against the separate allergens was far higher than that detected against the WME, thus indicating that the concentration of these can be limiting in the WME. We found no significant correlations between the RAST levels against Der p 2 and either Der p 5 or 7, and RAST-inhibition tests indicated a lack of cross-reactivity between Der p 2 and the other two allergens. In contrast, the RAST results revealed the existence of a significant immunological relationship between Der p 5 and 7. Although a certain degree of reactivity against the GST fusion partner was found in the allergic patients studied, this was not a significant influence in determining the positivity against the recombinant allergens. These results confirm the in vivo biological activity of recombinant Der p 2, 5 and 7, and indicate that whilst Der p 2 is undoubtedly a major mite allergen, both Der p 5 and 7 make important contributions toward the overall allergenic activity of house-dust mites.

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“…92 Der p 7-reactive IgE serum antibodies are found in only 53% of allergic patients, 93 however, the scale of the response can be as great as for the more major allergens. 94 )-though with expectedly low sequence identities (14% and 12%, respectively).…”

Section: Case Study #3: Der P 7 From Dermatophagoides Pteronyssinusmentioning

confidence: 99%

A synergistic approach to protein crystallization: Combination of a fixed‐arm carrier with surface entropy reduction

et al. 2010

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Protein crystallographers are often confronted with recalcitrant proteins not readily crystallizable, or which crystallize in problematic forms. A variety of techniques have been used to surmount such obstacles: crystallization using carrier proteins or antibody complexes, chemical modification, surface entropy reduction, proteolytic digestion, and additive screening. Here we present a synergistic approach for successful crystallization of proteins that do not form diffraction quality crystals using conventional methods. This approach combines favorable aspects of carrier-driven crystallization with surface entropy reduction. We have generated a series of maltose binding protein (MBP) fusion constructs containing different surface mutations designed to reduce surface entropy and encourage crystal lattice formation. The MBP advantageously increases protein expression and solubility, and provides a streamlined purification protocol. Using this technique, we have successfully solved the structures of three unrelated proteins that were previously unattainable. This crystallization technique represents a valuable rescue strategy for protein structure solution when conventional methods fail.

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Molecular cloning of a house dust mite allergen with common antibody binding specificities with multiple components in mite extracts

Cited by 57 publications

References 22 publications

Characterization and Immunobiology of House Dust Mite Allergens

Characterization and Immunobiology of House Dust Mite Allergens

Biological Activity of Recombinant Der p 2, Der p 5 and Der p 7 Allergens of the House-Dust Mite <i>Dermatophagoides pteronyssinus</i>

A synergistic approach to protein crystallization: Combination of a fixed‐arm carrier with surface entropy reduction

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