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Assignment of disulphide bridges in Par j 2.0101, a major allergen of Parietaria judaica pollen.
Biol Chem. 2003 Aug; 384(8):1165-72.BC

Abstract

Par j 2.0101, a major allergen of the Parietaria judaica pollen, was expressed in E. coli, purified to homogeneity and fully characterised both at the structural and the functional level. The recombinant rPar j 2.0101 protein showed an allergenic activity in histamine release, skin prick tests and capacity to bind IgE, almost identical to that of the native allergens purified from aqueous pollen extract. The complete pattern of S-S bridges of rPar j 2.0101 was determined by enzymatic digestion with endoproteinase Lys-C followed by mass spectrometric analysis of the resulting peptide mixtures. The eight cysteines occurring in the allergenic protein were found to be paired into the following four disulphides: Cys35-Cys83, Cys45-Cys60, Cys61-Cys106 and Cys81-Cys121. This structural information probes Par j 2.0101 to attain a 3-D fold consistent with that of the non-specific lipid transfer protein (ns-LTP) family and it represents an effective molecular basis to develop modified antigens by selective site-directed mutagenesis for immunotherapy.

Authors+Show Affiliations

Dipartimento di Chimica Organica e Biochimica, Università di Napoli Federico II, via Cinthia 6, 1-80126 Napoli, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12974385

Citation

Amoresano, Angela, et al. "Assignment of Disulphide Bridges in Par J 2.0101, a Major Allergen of Parietaria Judaica Pollen." Biological Chemistry, vol. 384, no. 8, 2003, pp. 1165-72.
Amoresano A, Pucci P, Duro G, et al. Assignment of disulphide bridges in Par j 2.0101, a major allergen of Parietaria judaica pollen. Biol Chem. 2003;384(8):1165-72.
Amoresano, A., Pucci, P., Duro, G., Colombo, P., Costa, M. A., Izzo, V., Lamba, D., & Geraci, D. (2003). Assignment of disulphide bridges in Par j 2.0101, a major allergen of Parietaria judaica pollen. Biological Chemistry, 384(8), 1165-72.
Amoresano A, et al. Assignment of Disulphide Bridges in Par J 2.0101, a Major Allergen of Parietaria Judaica Pollen. Biol Chem. 2003;384(8):1165-72. PubMed PMID: 12974385.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Assignment of disulphide bridges in Par j 2.0101, a major allergen of Parietaria judaica pollen. AU - Amoresano,Angela, AU - Pucci,Piero, AU - Duro,Giovanni, AU - Colombo,Paolo, AU - Costa,Maria A, AU - Izzo,Vincenzo, AU - Lamba,Doriano, AU - Geraci,Domenico, PY - 2003/9/17/pubmed PY - 2004/5/21/medline PY - 2003/9/17/entrez SP - 1165 EP - 72 JF - Biological chemistry JO - Biol Chem VL - 384 IS - 8 N2 - Par j 2.0101, a major allergen of the Parietaria judaica pollen, was expressed in E. coli, purified to homogeneity and fully characterised both at the structural and the functional level. The recombinant rPar j 2.0101 protein showed an allergenic activity in histamine release, skin prick tests and capacity to bind IgE, almost identical to that of the native allergens purified from aqueous pollen extract. The complete pattern of S-S bridges of rPar j 2.0101 was determined by enzymatic digestion with endoproteinase Lys-C followed by mass spectrometric analysis of the resulting peptide mixtures. The eight cysteines occurring in the allergenic protein were found to be paired into the following four disulphides: Cys35-Cys83, Cys45-Cys60, Cys61-Cys106 and Cys81-Cys121. This structural information probes Par j 2.0101 to attain a 3-D fold consistent with that of the non-specific lipid transfer protein (ns-LTP) family and it represents an effective molecular basis to develop modified antigens by selective site-directed mutagenesis for immunotherapy. SN - 1431-6730 UR - https://www.unboundmedicine.com/medline/citation/12974385/Assignment_of_disulphide_bridges_in_Par_j_2_0101_a_major_allergen_of_Parietaria_judaica_pollen_ L2 - https://www.degruyter.com/document/doi/10.1515/BC.2003.129 DB - PRIME DP - Unbound Medicine ER -