Molecular Characterization of Major Cat Allergen Fel d 1

Seppälä, Ulla; Hägglund, Per; Würtzen, Peter Adler; Ipsen, Henrik; Thorsted, Peter; Lenhard, Thomas; Roepstorff, Peter; Spangfort, Michael D.

doi:10.1074/jbc.m410668200

Cited by 23 publications

(12 citation statements)

References 38 publications

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“…The importance of Fel d 1 glycosylation in protein structure and immune response has also been somewhat controversial. While some studies show that different glycosylation patterns do not affect IgE production in vitro [ 9 , 14 , 15 ], a most recent study demonstrated that the mannose receptor has an essential role in internalizing Fel d 1. Mannose receptor cysteine rich domain recognizes the carbohydrates in Fel d 1 and in vivo assays showed that knockout mice for mannose receptor produced lower levels of immunoglobulins E and G [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

The Calcium Goes Meow: Effects of Ions and Glycosylation on Fel d 1, the Major Cat Allergen

et al. 2015

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The major cat allergen, Fel d 1, is a structurally complex protein with two N-glycosylation sites that may be filled by different glycoforms. In addition, the protein contains three putative Ca2+ binding sites. Since the impact of these Fel d 1 structure modifications on the protein dynamics, physiology and pathology are not well established, the present work employed computational biology techniques to tackle these issues. While conformational effects brought upon by glycosylation were identified, potentially involved in cavity volume regulation, our results indicate that only the central Ca2+ ion remains coordinated to Fel d 1 in biological solutions, impairing its proposed role in modulating phospholipase A2 activity. As these results increase our understanding of Fel d 1 structural biology, they may offer new support for understanding its physiological role and impact into cat-promoted allergy.

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Section: Introductionmentioning

confidence: 99%

The Calcium Goes Meow: Effects of Ions and Glycosylation on Fel d 1, the Major Cat Allergen

et al. 2015

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“…Both recombinant and naturally derived Fel d 1 have been used to investigate its IgE‐binding sites. Fel d 1 has been shown to be glycosylated and to undergo post‐translational modifications, both of which may have an effect on IgE recognition of conformational epitopes [7, 8]. Since the identification of the major IgE‐binding allergens in cat dander, T cell responses to both cat dander and Fel d 1 protein have been investigated.…”

Section: Introductionmentioning

confidence: 99%

Phenotypic analysis of perennial airborne allergen‐specific CD4⁺ T cells in atopic and non‐atopic individuals

Crack

Chan

McPherson

et al. 2011

Clin Experimental Allergy

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“…Reduction and alkylation of rVes v 2 were performed as described in Seppala et al (2005). Assignment of the disulfide bridges in r Ves v 2 and the glycosylation sites in nVes v 2 were performed by enzymatic digestion employing 3%(w/w) trypsin (Sequencing Grade Modified Trypsin, Promega, Madison, WI, USA) in 50 mM Tris-HCl pH 8.5, 0.2 M NaCl and 1 M urea.…”

Section: Characterization and Mass-spectrometric Analysismentioning

confidence: 99%

“…Natural Ves v 2 and rVes v 2 were redissolved in 50 mM Tris-HCl pH 8.5, 0.2 M NaCl and 1 M urea and incubated with 3%(w/w) endoproteinase Lys-C (Wako GmbH, Richmond, VA, USA) and Asp-N (Calbiochem, Sunnyvale, CA, USA) at 310 K for 18 h (Seppala et al, 2005). Reduction and alkylation of rVes v 2 were performed as described in Seppala et al (2005).…”

Section: Characterization and Mass-spectrometric Analysismentioning

confidence: 99%

Structure of recombinant Ves v 2 at 2.0 Å resolution: structural analysis of an allergenic hyaluronidase from wasp venom

Skov

Seppälä

Coen

et al. 2006

Acta Crystallogr D Biol Crystallogr

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Wasp venom from Vespula vulgaris contains three major allergens: Ves v 1, Ves v 2 and Ves v 5. Here, the cloning, expression, biochemical characterization and crystal structure determination of the hyaluronidase Ves v 2 from family 56 of the glycoside hydrolases are reported. The allergen was expressed in Escherichia coli as an insoluble protein and refolded and purified to obtain full enzymatic activity. Three N‐glycosylation sites at Asn79, Asn99 and Asn127 were identified in Ves v 2 from a natural source by enzymatic digestions combined with MALDI–TOF mass spectrometry. The crystal structure of recombinant Ves v 2 was determined at 2.0 Å resolution and reveals a central (β/α)7 core that is further stabilized by two disulfide bonds (Cys19–Cys308 and Cys185–Cys197). Based on sequence alignments and structural comparison with the honeybee allergen Api m 2, it is proposed that a conserved cavity near the active site is involved in binding of the substrate. Surface epitopes and putative glycosylation sites have been compared with those of two other major group 2 allergens from Apis mellifera (honeybee) and Dolichovespula maculata (white‐faced hornet). The analysis suggests that the harboured allergic IgE‐mediated cross‐reactivity between Ves v 2 and the allergen from D. maculata is much higher than that between Ves v 2 and the allergen from A. mellifera.

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Molecular Characterization of Major Cat Allergen Fel d 1

Cited by 23 publications

References 38 publications

The Calcium Goes Meow: Effects of Ions and Glycosylation on Fel d 1, the Major Cat Allergen

The Calcium Goes Meow: Effects of Ions and Glycosylation on Fel d 1, the Major Cat Allergen

Phenotypic analysis of perennial airborne allergen‐specific CD4⁺ T cells in atopic and non‐atopic individuals

Structure of recombinant Ves v 2 at 2.0 Å resolution: structural analysis of an allergenic hyaluronidase from wasp venom

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Molecular Characterization of Major Cat Allergen Fel d 1

Cited by 23 publications

References 38 publications

The Calcium Goes Meow: Effects of Ions and Glycosylation on Fel d 1, the Major Cat Allergen

The Calcium Goes Meow: Effects of Ions and Glycosylation on Fel d 1, the Major Cat Allergen

Phenotypic analysis of perennial airborne allergen‐specific CD4+ T cells in atopic and non‐atopic individuals

Structure of recombinant Ves v 2 at 2.0 Å resolution: structural analysis of an allergenic hyaluronidase from wasp venom

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Phenotypic analysis of perennial airborne allergen‐specific CD4⁺ T cells in atopic and non‐atopic individuals