Structural, Functional, and Immunological Characterization of Profilin Panallergens Amb a 8, Art v 4, and Bet v 2

Offermann, L.R.; Schlachter, Caleb R.; Perdue, Makenzie L.; Majorek, K.A.; He, John Z.; Booth, William T.; Garrett, Jessica; Kowal, Krzysztof; Chruszcz, M.

doi:10.1074/jbc.m116.733659

Cited by 38 publications

(42 citation statements)

References 72 publications

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“…Our results on the thermal stabilities of Amb a 8, Art v 4, and Bet v 2 (inhaled allergens originating from pollens) clearly show that indeed these proteins are relatively unstable. 33 The presence of the His-tag has no significant impact on the thermal stabilities of Amb a 8 and Art v 4, but it slightly increases the stability of Bet v 2. This observation is consistent with the high sequence identity between Amb a 8 and Art v 4 (∼89%) and lower sequence identity between Bet v 2 and the two other profilins (∼76%).…”

Section: Resultsmentioning

confidence: 98%

Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability

et al. 2018

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For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant proteins, a consequence of His-tag addition is that it can affect protein function and stability. Functional proteins are essential in the elucidation of their biological, kinetic, structural, and thermodynamic properties. In this study, we determine the effect of N-terminal His-tags on the thermal stability of select proteins using differential scanning fluorimetry and identify that the removal of the His-tag can have both beneficial and deleterious effects on their stability.

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Section: Resultsmentioning

confidence: 98%

Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability

et al. 2018

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“…These differences were also manifested in solution by a significantly higher binding affinity of the oxidized (disulfide-bonded) than the reduced form towards poly- l -proline resin, as evidenced by a preferential elution of the reduced Bet v 2 form. Recently, a crystal structure of the complex of latex profilin with poly- l -proline has revealed the direct involvement of the terminal helices in ligand binding [ 10 ]. The poly- l -proline binding sites of Bet v 2 map onto the immediate neighborhood of the Cys13-Cys117 disulfide bond, providing a direct link between the differences in proline binding with the differences in the crystal structures of both Bet v 2 forms ( Figure 5 ).…”

Section: Discussionmentioning

confidence: 99%

“…A recent study showed that rubber tree profilin, Hev b 8, was able to homodimerize via an inter-molecular disulfide bridge [ 9 ]. Another study also observed an intra-disulfide bond between Cys95 and Cys117 in the three cysteines containing Art v 4 crystal structure, however, it is not observed in all the molecules [ 10 ]. It needs to be pointed out that the cysteine 95 is not conserved in profilin.…”

Section: Introductionmentioning

confidence: 99%

Two Distinct Conformations in Bet v 2 Determine Its Proteolytic Resistance to Cathepsin S

Soh

Briza

Dall

et al. 2017

IJMS

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Birch pollen allergy affects more than 20% of the European allergic population. On a molecular level, birch pollen allergy can be linked to the two dominant allergens Bet v 1 and Bet v 2. Bet v 2 belongs to the profilin family, which is abundant in the plant kingdom. Importantly, the homologous plant profilins have a conserved cysteine motif with a currently unknown functional relevance. In particular, it is unknown whether the motif is relevant for disulfide formation and to what extent it would affect the profilins’ structural, functional and immunological properties. Here we present crystal structures of Bet v 2 in the reduced and the oxidized state, i.e., without and with a disulfide bridge. Despite overall structural similarity, the two structures distinctly differ at their termini which are stabilized to each other in the oxidized, i.e., disulfide-linked state. These structural differences translate into differences in their proteolytic resistance. Whereas the oxidized Bet v 2 is rather resistant towards the endolysosomal protease cathepsin S, it is rapidly degraded in the reduced form. By contrast, both Bet v 2 forms exhibit similar immunological properties as evidenced by their binding to IgE antibodies from birch pollen allergic patients and by their ability to trigger histamine release in a humanized rat basophilic leukemia cells (RBL) assay, independent of the presence or absence of the disulfide bridge. Taken together our findings suggest that the oxidized Bet v 2 conformation should be the relevant species, with a much longer retention time to trigger immune responses.

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“…Table S1c). In addition, Amb a 8.0101 shares 89.5% of its AA sequence with Art v 4.0101, displaying comparable cross-reactivity within the same family due to structural similarities between the 2 proteins [61]. There is a close relationship between ragweed and sunflower proteins, as the highest homology outside the species is recorded between ragweed and sunflower, mainly for profilin (94% sequence homology, see online suppl.…”

Section: Minor Ragweed Pollen Allergensmentioning

confidence: 99%

“…Further characterization revealed the structure showing 2 terminal alpha-helices, 1 short alpha-helix, and a beta-hairpin that sandwich a central 5-stranded antiparallel beta-sheet [61] (Fig. 4).…”

Section: Minor Ragweed Pollen Allergensmentioning

confidence: 99%

Ragweed Pollen Allergy: Burden, Characteristics, and Management of an Imported Allergen Source in Europe

Chen

Marusciac

Tamas

et al. 2018

Int Arch Allergy Immunol

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Ambrosia artemisiifolia, also known as common or short ragweed, is an invasive annual flowering herbaceous plant that has its origin in North America. Nowadays, ragweed can be found in many areas worldwide. Ragweed pollen is known for its high potential to cause type I allergic reactions in late summer and autumn and represents a major health problem in America and several countries in Europe. Climate change and urbanization, as well as long distance transport capacity, enhance the spread of ragweed pollen. Therefore ragweed is becoming domestic in non-invaded areas which in turn will increase the sensitization rate. So far 11 ragweed allergens have been described and, according to IgE reactivity, Amb a 1 and Amb a 11 seem to be major allergens. Sensitization rates of the other allergens vary between 10 and 50%. Most of the allergens have already been recombinantly produced, but most of them have not been characterized regarding their allergenic activity, therefore no conclusion on the clinical relevance of all the allergens can be made, which is important and necessary for an accurate diagnosis. Pharmacotherapy is the most common treatment for ragweed pollen allergy but fails to impact on the course of allergy. Allergen-specific immunotherapy (AIT) is the only causative and disease-modifying treatment of allergy with long-lasting effects, but currently it is based on the administration of ragweed pollen extract or Amb a 1 only. In order to improve ragweed pollen AIT, new strategies are required with higher efficacy and safety.

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Structural, Functional, and Immunological Characterization of Profilin Panallergens Amb a 8, Art v 4, and Bet v 2

Cited by 38 publications

References 72 publications

Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability

Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability

Two Distinct Conformations in Bet v 2 Determine Its Proteolytic Resistance to Cathepsin S

Ragweed Pollen Allergy: Burden, Characteristics, and Management of an Imported Allergen Source in Europe

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