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Detection and localisation of disulphide bonds in a synthetic peptide reproducing the sequence 1–30 of Par j 1.0101 by electrospray ionisation mass spectrometry

✍ Scribed by Vincenzo Cunsolo; Salvatore Foti; Rosaria Saletti; Leopoldo Ceraulo; Vita Di Stefano


Publisher
John Wiley and Sons
Year
2001
Tongue
English
Weight
79 KB
Volume
1
Category
Article
ISSN
1615-9853

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✦ Synopsis


Detection and localisation of disulphide bonds in a synthetic peptide reproducing the sequence 1-30 of Par j 1.0101 by electrospray ionisation mass spectrometry

The structural characterisation of a synthetic peptide reproducing the sequence 1-30 of Par j 1.0101, a major allergenic protein present in the pollen of Parietaria judaica, by combined use of chemical and enzymatic cleavage, reversed-phase highperformance liquid chromatography and electrospray ionisation mass spectrometry (ESI-MS), is described. Direct ESI-MS of the synthetic peptide after reaction with methyl iodide showed that the product is a mixture of two peptides: one form in which two out of the four cysteine residues present in the sequence are oxidised and a minor amount of another form in which all the cysteines are fully reduced. It was ascertained, using the combined procedure indicated above and without prior separation of the two species, that the disulphide bond in the partially oxidised form is located between cysteines 29 and 30. These results show the usefulness of this approach for characterising synthetic peptides containing multiple cysteine residues in the sequence.