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Publications (10 of 53) Show all publications
Nilsson, O. B., Adedoyin, J., Rhyner, C., Neimert-Andersson, T., Grundström, J., Berndt, K. D., . . . Grönlund, H. (2011). In vitro evolution of allergy vaccine candidates, with maintained structure, but reduced B cell and T cell activation capacity. PLoS ONE, 6, Article ID e24558.
Open this publication in new window or tab >>In vitro evolution of allergy vaccine candidates, with maintained structure, but reduced B cell and T cell activation capacity
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2011 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 6, article id e24558Article in journal (Refereed) Published
Abstract [en]

Allergy and asthma to cat (Felis domesticus) affects about 10% of the population in affluent countries. Immediate allergic symptoms are primarily mediated via IgE antibodies binding to B cell epitopes, whereas late phase inflammatory reactions are mediated via activated T cell recognition of allergen-specific T cell epitopes. Allergen-specific immunotherapy relieves symptoms and is the only treatment inducing a long-lasting protection by induction of protective immune responses. The aim of this study was to produce an allergy vaccine designed with the combined features of attenuated T cell activation, reduced anaphylactic properties, retained molecular integrity and induction of efficient IgE blocking IgG antibodies for safer and efficacious treatment of patients with allergy and asthma to cat. The template gene coding for rFel d 1 was used to introduce random mutations, which was subsequently expressed in large phage libraries. Despite accumulated mutations by up to 7 rounds of iterative error-prone PCR and biopanning, surface topology and structure was essentially maintained using IgE-antibodies from cat allergic patients for phage enrichment. Four candidates were isolated, displaying similar or lower IgE binding, reduced anaphylactic activity as measured by their capacity to induce basophil degranulation and, importantly, a significantly lower T cell reactivity in lymphoproliferative assays compared to the original rFel d 1. In addition, all mutants showed ability to induce blocking antibodies in immunized mice.The approach presented here provides a straightforward procedure to generate a novel type of allergy vaccines for safer and efficacious treatment of allergic patients.

National Category
Structural Biology
Identifiers
urn:nbn:se:sh:diva-29034 (URN)10.1371/journal.pone.0024558 (DOI)000295321800032 ()21931754 (PubMedID)2-s2.0-80052768250 (Scopus ID)
Available from: 2016-01-07 Created: 2016-01-07 Last updated: 2017-12-01Bibliographically approved
Elgán, T. H., Planson, A.-G. -., Beckwith, J., Güntert, P. & Berndt, K. D. (2010). Determinants of activity in glutaredoxins: An in vitro evolved Grx1-like variant of Escherichia coli Grx3. Biochemical Journal, 430(3), 487-495
Open this publication in new window or tab >>Determinants of activity in glutaredoxins: An in vitro evolved Grx1-like variant of Escherichia coli Grx3
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2010 (English)In: Biochemical Journal, ISSN 0264-6021, E-ISSN 1470-8728, Vol. 430, no 3, p. 487-495Article in journal (Refereed) Published
Abstract [en]

The Escherichia coli glutaredoxins 1 and 3 (Grx1 and Grx3) are structurally similar (37% sequence identity), yet have different activities in vivo. Unlike Grx3, Grx1 efficiently reduces protein disulfides in proteins such as RR (ribonucleotide reductase), whereas it is poor at reducing S-glutathionylated proteins. An E. coli strain lacking genes encoding thioredoxins 1 and 2 and Grx1 is not viable on either rich or minimal medium; however, a M43V mutation in Grx3 restores growth under these conditions and results in a Grx1-like protein [Ortenberg, Gon, Porat and Beckwith (2004) Proc. Natl. Acad. Sci. U.S.A. 101, 7439-7944]. To uncover the structural basis of this change in activity, we have compared wild-type and mutant Grx3 using CD and NMR spectroscopy. Ligand-induced stability measurements demonstrate that the Grx3(M43V/C65Y) mutant has acquired affinity for RR. Far-UV CD spectra reveal no significant differences, but differences are observed in the near-UV region indicative of tertiary structural changes. NMR 1H- 15N HSQC (heteronuclear single quantum coherence) spectra show that approximately half of the 82 residues experience significant (Δδ > 0.03 p.p.m.) chemical shift deviations in the mutant, including nine residues experiencing extensive (Δδ ≥ 0.15 p.p.m.) deviations. To test whether the M43V mutation alters dynamic properties of Grx3, H/D (hydrogen/deuterium) exchange experiments were performed demonstrating that the rate at which backbone amides exchange protons with the solvent is dramatically enhanced in the mutant, particularly in the core of the protein. These data suggest that the Grx1-like activity of the Grx3(M43V/C65Y) mutant may be explained by enhanced intrinsic motion allowing for increased specificity towards larger substrates such as RR.

Keywords
Chemical shift, Glutaredoxin (Grx), Hydrogen/deuterium exchange, Ligand-induced stability, NMR, Ribonucleotide reductase, CD spectra, Dynamic property, E. coli, Genes encoding, Glutaredoxin, Heteronuclear single-quantum coherences, In-vitro, In-vivo, Intrinsic motions, Larger substrates, NMR spectroscopy, Sequence identity, Stability measurements, Structural basis, Structural change, Thioredoxins, UV region, Wild types, Amides, Enzymes, Escherichia coli, Ligands, Nuclear magnetic resonance spectroscopy, Quantum theory, Chemical stability, deuterium, glutaredoxin 1, glutaredoxin 3, hydrogen, unclassified drug, article, binding affinity, circular dichroism, controlled study, enzyme activation, gene mutation, heteronuclear single quantum coherence, in vitro study, nonhuman, priority journal, protein structure, protein tertiary structure, proton nuclear magnetic resonance, ultraviolet radiation, wild type, Algorithms, Amino Acid Substitution, Enzyme Stability, Escherichia coli Proteins, Glutaredoxins, Glutathione Disulfide, Isoenzymes, Magnetic Resonance Spectroscopy, Models, Molecular, Mutant Proteins, Mutation, Oxidation-Reduction, Protein Conformation, Protein Structure, Tertiary, Thermodynamics
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:sh:diva-20777 (URN)10.1042/BJ20100289 (DOI)000282246700011 ()20604742 (PubMedID)2-s2.0-77956697929 (Scopus ID)
Available from: 2013-12-18 Created: 2013-12-18 Last updated: 2017-12-06Bibliographically approved
Ferreira, M. E., Berndt, K. D., Nilsson, J. & Wright, A. P. H. (2010). WD40 Domain Divergence Is Important for Functional Differences between he Fission Yeast Tup11 and Tup12 Co-Repressor Proteins. PLoS ONE, 5(6), e11009
Open this publication in new window or tab >>WD40 Domain Divergence Is Important for Functional Differences between he Fission Yeast Tup11 and Tup12 Co-Repressor Proteins
2010 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 5, no 6, p. e11009-Article in journal (Refereed) Published
Abstract [en]

We have previously demonstrated that subsets of Ssn6/Tup target genes ave distinct requirements for the Schizosaccharomyces pombe homologs of he Tup1/Groucho/TLE co-repressor proteins, Tup11 and Tup12. The very igh level of divergence in the histone interacting repression domains f the two proteins suggested that determinants distinguishing Tup11 and up12 might be located in this domain. Here we have combined hylogenetic and structural analysis as well as phenotypic haracterization, under stress conditions that specifically require up12, to identify and characterize the domains involved in up12-specific action. The results indicate that divergence in the epression domain is not generally relevant for Tup12-specific function. nstead, we show that the more highly conserved C-terminal WD40 repeat omain of Tup12 is important for Tup12-specific function. Surface amino cid residues specific for the WD40 repeat domain of Tup12 proteins in ifferent fission yeasts are clustered in blade 3 of the propeller-like tructure that is characteristic of WD40 repeat domains. The Tup11 and up12 proteins in fission yeasts thus provide an excellent model system or studying the functional divergence of WD40 repeat domains.

National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:sh:diva-13706 (URN)10.1371/journal.pone.0011009 (DOI)000278494900021 ()2-s2.0-77955193861 (Scopus ID)
Available from: 2011-12-06 Created: 2011-12-06 Last updated: 2017-12-08Bibliographically approved
Ferreira, M. E., Prochasson, P., Berndt, K. D., Workman, J. L. & Wright, A. P. H. (2009). Activator-binding domains of the SWI/SNF chromatin remodeling complex characterized in vitro are required for its recruitment to promoters in vivo. The FEBS Journal, 276(9), 2557-2565
Open this publication in new window or tab >>Activator-binding domains of the SWI/SNF chromatin remodeling complex characterized in vitro are required for its recruitment to promoters in vivo
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2009 (English)In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658, Vol. 276, no 9, p. 2557-2565Article in journal (Refereed) Published
Abstract [en]

Interaction between acidic activation domains and the activator-binding domains of Swi1 and Snf5 of the yeast SWI/SNF chromatin remodeling complex has previously been characterized in vitro. Although deletion of both activator-binding domains leads to phenotypes that differ from the wild-type, their relative importance for SWI/SNF recruitment to target genes has not been investigated. In the present study, we used chromatin immunoprecipitation assays to investigate the individual and collective importance of the activator-binding domains for SWI/SNF recruitment to genes within the GAL regulon in vivo. We also investigated the consequences of defective SWI/SNF recruitment for target gene activation. We demonstrate that deletion of both activator-binding domains essentially abolishes galactose-induced SWI/SNF recruitment and causes a reduction in transcriptional activation similar in magnitude to that associated with a complete loss of SWI/SNF activity. The activator-binding domains in Swi1 and Snf5 make approximately equal contributions to the recruitment of SWI/SNF to each of the genes studied. The requirement for SWI/SNF recruitment correlates with GAL genes that are highly and rapidly induced by galactose.

National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:sh:diva-13900 (URN)10.1111/j.1742-4658.2009.06979.x (DOI)000264882700012 ()19476494 (PubMedID)2-s2.0-64149100814 (Scopus ID)
Available from: 2011-12-14 Created: 2011-12-14 Last updated: 2017-12-08Bibliographically approved
Vilhelmsson, M., Glaser, A. G., Martinez, D. B., Schmidt, M., Johansson, C., Rhyner, C., . . . Zargari, A. (2008). Mutational analysis of amino acid residues involved in IgE-binding to the Malassezia sympodialis allergen Mala s 11. Molecular Immunology, 46(2), 294-303
Open this publication in new window or tab >>Mutational analysis of amino acid residues involved in IgE-binding to the Malassezia sympodialis allergen Mala s 11
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2008 (English)In: Molecular Immunology, ISSN 0161-5890, E-ISSN 1872-9142, Vol. 46, no 2, p. 294-303Article in journal (Refereed) Published
Abstract [en]

The yeast Malassezia sympodialis, which is an integral part of the normal cutaneous flora, has been shown to elicit specific IgE- and T-cell reactivity in atopic eczema (AE) patients. The M. sympodialis allergen Mala s 11 has a high degree of amino acid sequence homology to manganese superoxide dismutase (MnSOD) from Homo sapiens (50%) and Aspergillus fumigatus (56%). Humoral and cell-mediated cross-reactivity between MnSOD from H. sapiens and A. fumigatus has been demonstrated. Taken together with the recent finding that human MnSOD (hMnSOD) can act as an autoallergen in AE patients sensitised to M. sympodialis, we hypothesized that cross-reactivity could also occur between hMnSOD and Mala s 11, endogenous hMnSOD thus being capable of stimulating an immune response through molecular mimicry. Herein we demonstrate that recombinant Mala s 11 (rMala s 11) is able to inhibit IgE-binding to recombinant hMnSOD and vice versa, indicating that these two homologues share common IgE-binding epitopes and providing an explanation at a molecular level for the autoreactivity to hMnSOD observed in AE patients sensitised to Mala s 11. Using molecular modelling and mapping of identical amino acids exposed on the surface of both Mala s 11 and hMnSCE) we identified four regions each composed of 4-5 residues which are potentially involved in IgE-mediated cross-reactivity. Mutated rMala s 11 molecules were produced in which these residues were altered. Native-like folding was verified by enzymatic activity tests and circular dichroism. The rMala s 11 mutants displayed lower IgE-binding in comparison to wild-type rMala s 11 using plasma from AE patients. In particular, mutation of the residues E29, P30, E122 and K125 lowered the IgE-binding to Mala s 11. The results of this study provide new insights in the molecular basis underlying the cross-reactivity between Mala s 11 and hMnSOD.

National Category
Biochemistry and Molecular Biology Immunology
Identifiers
urn:nbn:se:sh:diva-14112 (URN)10.1016/j.molimm.2008.07.036 (DOI)000261296200009 ()18922581 (PubMedID)2-s2.0-54849437704 (Scopus ID)
Available from: 2011-12-19 Created: 2011-12-16 Last updated: 2017-12-08Bibliographically approved
Elgan, T. H. & Berndt, K. D. (2008). Quantifying Escherichia coli Glutaredoxin-3 Substrate Specificity Using Ligand-induced Stability. Journal of Biological Chemistry, 283(47), 32839-32847
Open this publication in new window or tab >>Quantifying Escherichia coli Glutaredoxin-3 Substrate Specificity Using Ligand-induced Stability
2008 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 283, no 47, p. 32839-32847Article in journal (Refereed) Published
Abstract [en]

Traditionally, quantification of protein-ligand affinity is performed using kinetic or equilibrium measurements. However, if the binding reaction proceeds via a stable covalent complex, these approaches are often limited. By exploiting the fact that the conformational stabilization of a protein is altered upon ligand binding due to specific interactions, and using an array of selectively chosen ligand analogs, one can quantify the contribution individual interactions have on specificity. We have used ligand-induced stability as a basis to dissect the interaction between glutaredoxin-3 (Grx3) and one of its native substrates, the tripeptide glutathione. Taking advantage of the fact that Grx3 can be trapped in a covalent mixed disulfide to glutathione or to selected synthetic glutathione analogs as part of the natural catalytic cycle, individual contributions to binding of specific molecular groups can be quantified by changes in ligand-induced stability. These changes in conformational stability are interpreted in terms of interaction energies (i.e. specificity) of the particular groups present on the ligand analog. Our results illustrate that although Grx3 recognizes glutathione predominantly through independent and additive ionic interactions at the N- and C-terminal of glutathione, van der Waals interactions from the unique gamma-glutamate moiety of glutathione also play an important role. This study places us closer to understanding the complex task of accommodating multiple substrate specificities in proteins of the thioredoxin superfamily and underscores the general applicability of ligand-induced stability to probe substrate specificity.

National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:sh:diva-22529 (URN)10.1074/jbc.M804019200 (DOI)000260893700073 ()18757366 (PubMedID)2-s2.0-57749093461 (Scopus ID)
Available from: 2014-03-03 Created: 2014-03-03 Last updated: 2017-12-05Bibliographically approved
Lundholm, L., Bryzgalova, G., Gao, H., Portwood, N., Fält, S., Berndt, K. D., . . . Khan, A. (2008). The estrogen receptor {alpha}-selective agonist propyl pyrazole triol improves glucose tolerance in ob/ob mice; potential molecular mechanisms. Journal of Endocrinology, 199(2), 275-286
Open this publication in new window or tab >>The estrogen receptor {alpha}-selective agonist propyl pyrazole triol improves glucose tolerance in ob/ob mice; potential molecular mechanisms
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2008 (English)In: Journal of Endocrinology, ISSN 0022-0795, E-ISSN 1479-6805, Vol. 199, no 2, p. 275-286Article in journal (Refereed) Published
Abstract [en]

The aim of this study was to validate the role of estrogen receptor alpha (ERalpha) signaling in the regulation of glucose metabolism, and to compare the molecular events upon treatment with the ERalpha-selective agonist propyl pyrazole triol (PPT) or 17beta-estradiol (E(2)) in ob/ob mice. Female ob/ob mice were treated with PPT, E(2) or vehicle for 7 or 30 days. Intraperitoneal glucose and insulin tolerance tests were performed, and insulin secretion was determined from isolated islets. Glucose uptake was assayed in isolated skeletal muscle and adipocytes. Gene expression profiling in the liver was performed using Affymetrix microarrays, and the expression of selected genes was studied by real-time PCR analysis. PPT and E(2) treatment improved glucose tolerance and insulin sensitivity. Fasting blood glucose levels decreased after 30 days of PPT and E(2) treatment. However, PPT and E(2) had no effect on insulin secretion from isolated islets. Basal and insulin-stimulated glucose uptake in skeletal muscle and adipose tissue were similar in PPT and vehicle-treated ob/ob mice. Hepatic lipid content was decreased after E(2) treatment. In the liver, treatment with E(2) and PPT increased and decreased the respective expression levels of the transcription factor signal transducer and activator of transcription 3, and of glucose-6-phosphatase. In summary, our data demonstrate that PPT exerts anti-diabetic effects, and these effects are mediated via ERalpha.

National Category
Cell Biology
Identifiers
urn:nbn:se:sh:diva-29031 (URN)10.1677/JOE-08-0192 (DOI)000260768600013 ()18757549 (PubMedID)
Note

Lundholm, L Bryzgalova, G Gao, H Portwood, N Falt, S Berndt, K D Dicker, A Galuska, D Zierath, J R Gustafsson, J-A Efendic, S Dahlman-Wright, K Khan, A Research Support, Non-U.S. Gov't England The Journal of endocrinology J Endocrinol. 2008 Nov;199(2):275-86. Epub 2008 Aug 29.

Available from: 2016-01-07 Created: 2016-01-07 Last updated: 2017-12-01Bibliographically approved
Wu, X., Oppermann, M., Berndt, K. D., Bergman, T., Jörnvall, H., Knapp, S. & Oppermann, U. (2008). Thermal unfolding of the archaeal DNA and RNA binding protein Ssh10. Biochemical and Biophysical Research Communications - BBRC, 373(4), 482-487
Open this publication in new window or tab >>Thermal unfolding of the archaeal DNA and RNA binding protein Ssh10
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2008 (English)In: Biochemical and Biophysical Research Communications - BBRC, ISSN 0006-291X, E-ISSN 1090-2104, Vol. 373, no 4, p. 482-487Article in journal (Refereed) Published
Abstract [en]

The reversible thermal unfolding of the archaeal histone-like protein Ssh10b from the extremophile Sulfolobus shibatae was studied using differential scanning calorimetry and circular dichroism spectroscopy. Analytical ultracentrifugation and gel filtration showed that Ssh10b is a stable dimer in the pH range 2.5-7.0. Thermal denaturation data fit into a two-state unfolding model, suggesting that the Ssh10 dimer unfolds as a single cooperative unit with a maximal melting temperature of 99.9 degrees C and an enthalpy change of 134 kcal/mol at pH 7.0. The heat capacity change upon unfolding determined from linear fits of the temperature dependence of DeltaH(cal) is 2.55 kcal/(mol K). The low specific heat capacity change of 13 cal/(mol K residue) leads to a considerable flattening of the protein stability curve (DeltaG (T)) and results in a maximal DeltaG of only 9.5 kcal/mol at 320 K and a DeltaG of only 6.0 kcal/mol at the optimal growth temperature of Sulfolobus.

Keywords
*Hot Temperature, Amino Acid Sequence, Archaeal Proteins/chemistry/*metabolism, Calorimetry, Differential Scanning, Circular Dichroism, DNA-Binding Proteins/chemistry/*metabolism, Histones/chemistry/*metabolism, Hydrogen-Ion Concentration, Molecular Sequence Data, Protein Denaturation, Protein Folding, RNA-Binding Proteins/chemistry/*metabolism, Sulfolobus/*metabolism, Thermodynamics
National Category
Biochemistry and Molecular Biology
Research subject
Other research area
Identifiers
urn:nbn:se:sh:diva-29045 (URN)10.1016/j.bbrc.2008.06.030 (DOI)000258208500006 ()18571501 (PubMedID)2-s2.0-48949115342 (Scopus ID)
Note

Wu, Xiaoqiu Oppermann, Madalina Berndt, Kurt D Bergman, Tomas Jornvall, Hans Knapp, Stefan Oppermann, Udo Research Support, Non-U.S. Gov't United States Biochemical and biophysical research communications Biochem Biophys Res Commun. 2008 Sep 5;373(4):482-7. Epub 2008 Jun 20.

Available from: 2016-01-07 Created: 2016-01-07 Last updated: 2017-12-01Bibliographically approved
Sagemark, J., Elgan, T. H., Bürglin, T. R., Johansson, C., Holmgren, A. & Berndt, K. D. (2007). Redox properties and evolution of human glutaredoxins. Proteins: Structure, Function, and Bioinformatics, 68(4), 879-892
Open this publication in new window or tab >>Redox properties and evolution of human glutaredoxins
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2007 (English)In: Proteins: Structure, Function, and Bioinformatics, ISSN 0887-3585, E-ISSN 1097-0134, Vol. 68, no 4, p. 879-892Article in journal (Refereed) Published
Abstract [en]

Glutaredoxins (Grxs) are glutathione-dependent oxidoreductases that belong to the thioredoxin superfamily catalyzing thiol-disulfide exchange reactions via active site cysteine residues. Focusing on the human dithiol glutaredoxins having a C-X-Y-C active site sequence motif, the redox potentials of hGrxl and hGrx2 were determined to be -232 and -221 mV, respectively, using a combination of redox buffers, protein-protein equilibrium and thermodynamic linkage. In addition, a nonactive site disulfide was identified between Cys28 and Cys.113 in hGrx2 using redox buffers and chemical digestion. This disulfide confers nearly five kcal mol-1 additional stability by linking the C-terminal helix to the bulk of the protein. The redox potential of this nonactive site disulfide was determined to be -317 mVand is thus expected to be present in all but the most reducing conditions in vivo. As all human glutaredoxins contain additional nonactive site cysteine residues, a full phylogenetic analysis was performed to help elucidate their structural and functional roles. Three distinct groups were found: Grx1, Grx2, and Grx5, the latter representing a highly conserved group of monothiol glutaredoxins having a C-G-F-S active site sequence, with clear homologs from bacteria to human. Grx1 and Grx2 diverged from a common ancestor before the origin vertebrates, possibly even earlier in animal evolution. The highly stabilizing nonactive site disulfide observed in hGrx2 is found to be a conserved feature within the deuterostomes and appears to be the only additional conserved intramolecular disulfide within the glutaredoxins.

National Category
Biochemistry and Molecular Biology Biophysics
Identifiers
urn:nbn:se:sh:diva-14208 (URN)10.1002/prot.21416 (DOI)000249188900008 ()17546662 (PubMedID)2-s2.0-34548836048 (Scopus ID)
Available from: 2011-12-20 Created: 2011-12-19 Last updated: 2018-01-26Bibliographically approved
Kaiser, L., Velickovic, T. C., Badia-Martinez, D., Adedoyin, J., Thunberg, S., Hallen, D., . . . Achour, A. (2007). Structural characterization of the tetrameric form of the major cat allergen Fel d 1. Journal of Molecular Biology, 370(4), 714-727
Open this publication in new window or tab >>Structural characterization of the tetrameric form of the major cat allergen Fel d 1
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2007 (English)In: Journal of Molecular Biology, ISSN 0022-2836, E-ISSN 1089-8638, Vol. 370, no 4, p. 714-727Article in journal (Refereed) Published
Abstract [en]

Felis domesticus allergen 1(Fel d 1) is a 35 kDa tetrameric glycoprotein formed by two heterodimers which elicits IgE responses in 95% of patients with allergy to cat. We have previously established in vitro conditions for the appropriate folding of recombinant Fel d 1 using a direct linkage of chain 1 to chain 2 (construct Fel d 1 (1+2)) and chain 2 to chain 1 (construct Fel d 1 (2+1)). Although the crystal structure of Fel d 1 (2+1) revealed a striking structural similarity to that of uteroglobin, a steroid-inducible cytokine-like molecule with anti-inflammatory and immunomodulatory properties, no functional tetrameric form of Fel d 1 could be identified. Here we present the crystal structure of the Fel d 1 (1+2) tetramer at 1.6 A resolution. Interestingly, the crystal structure of tetrameric Fel d 1 reveals two different calcium-binding sites. Symmetrically positioned on each side of the Fel d 1 tetramer, the external Ca(2+)-binding sites correspond to a putative Ca(2+)-binding site previously suggested for uteroglobin. The second Ca(2+)-binding site lies within the dimerization interface, stabilizing the formation of the Fel d 1 tetramer, and inducing important local conformational changes that directly govern the shape of two water-filled cavities. The crystal structure suggests a potential portal for an unknown ligand. Alternatively, the two cavities could be used by the allergen as a conditional inner space allowing for the spatial rearrangement of centrally localized side-chains, such as Asp130, without altering the overall fold of the molecule. The striking structural similarity of the major cat allergen to uteroglobin, coupled to the identification in the present study of a common Ca(2+)-binding site, let us speculate that Fel d 1 could provoke an allergic response through the modulation of phospholipase A2, by sequestering Ca ions in a similar manner as previously suggested for uteroglobin.

Keywords
Allergens/*chemistry/genetics/immunology/metabolis, Amino Acid Sequence, Animals, Binding Sites, Calcium/chemistry/metabolism, Cats/genetics/*immunology, Cell Proliferation, Cells, Cultured, Crystallography, X-Ray, Dimerization, Glycoproteins/*chemistry/genetics/immunology/metab, Hydrophobicity, Models, Molecular, Molecular Sequence Data, Protein Binding, Protein Structure, Quaternary, Uteroglobin/chemistry/metabolism
National Category
Structural Biology
Identifiers
urn:nbn:se:sh:diva-29024 (URN)10.1016/j.jmb.2007.04.074 (DOI)000247904400009 ()17543334 (PubMedID)
Note

Kaiser, Liselotte Velickovic, Tanja Cirkovic Badia-Martinez, Daniel Adedoyin, Justus Thunberg, Sarah Hallen, Dan Berndt, Kurt Gronlund, Hans Gafvelin, Guro van Hage, Marianne Achour, Adnane Research Support, Non-U.S. Gov't England Journal of molecular biology J Mol Biol. 2007 Jul 20;370(4):714-27. Epub 2007 May 10.

Available from: 2016-01-07 Created: 2016-01-07 Last updated: 2017-12-01Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-3049-967X

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