caa a22 4500 378853155 CHVBK 20180305123328.0 cr unu---uuuuu 161128e20030820xx s 000 0 eng 10.1515/BC.2003.129 doi (NATIONALLICENCE)gruyter-10.1515/BC.2003.129 Assignment of Disulphide Bridges in Par j 2.0101, a Major Allergen of Parietaria judaica Pollen [Elektronische Daten] [A. Amoresano, P. Pucci, G. Duro, P. Colombo, M. A. Costa, V. Izzo, D. Lamba, D. Geraci] 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 nonspecific lipid transfer protein (ns-LTP) family and it represents an effective molecular basis to develop modified antigens by selective site-directed mutagenesis for immunotherapy. Copyright © 2003 by Walter de Gruyter GmbH & Co. KG Biochemistry nationallicence Molecular biology nationallicence Cellular biology nationallicence Amoresano A. aut Pucci P. aut Duro G. aut Colombo P. aut Costa M. A. aut Izzo V. aut Lamba D. aut Geraci D. aut Biological Chemistry Walter de Gruyter 384/8(2003-08-20), 1165-1172 1431-6730 384:8<1165 2003 384 bchm https://doi.org/10.1515/BC.2003.129 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/BC.2003.129 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Amoresano A. aut NATIONALLICENCE 700 1- Pucci P. aut NATIONALLICENCE 700 1- Duro G. aut NATIONALLICENCE 700 1- Colombo P. aut NATIONALLICENCE 700 1- Costa M. A. aut NATIONALLICENCE 700 1- Izzo V. aut NATIONALLICENCE 700 1- Lamba D. aut NATIONALLICENCE 700 1- Geraci D. aut NATIONALLICENCE 773 0- Biological Chemistry Walter de Gruyter 384/8(2003-08-20), 1165-1172 1431-6730 384:8<1165 2003 384 bchm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter