Purification and structural stability of the peach allergens Pru p 1 and Pru p 3

A - Papers appearing in refereed journals

Gaier, S., Marsh, J., Oberhuber, C., Rigby, N. M., Lovegrove, A., Alessandri, S., Briza, P., Radauer, C., Zuidmeer, L., Van Ree, R., Hemmer, W., Sancho, A. I., Mills, C., Hoffmann-Sommergruber, K. and Shewry, P. R. 2008. Purification and structural stability of the peach allergens Pru p 1 and Pru p 3. Molecular Nutrition & Food Research. 52, pp. S220-S229.

AuthorsGaier, S., Marsh, J., Oberhuber, C., Rigby, N. M., Lovegrove, A., Alessandri, S., Briza, P., Radauer, C., Zuidmeer, L., Van Ree, R., Hemmer, W., Sancho, A. I., Mills, C., Hoffmann-Sommergruber, K. and Shewry, P. R.
Abstract

Pru p 1 (a Bet v 1 homologue) and Pru p 3 (a nonspecific lipid transfer protein; nsLTP) are major allergenic proteins in peach fruit, but differ in their abundance and stability. Pru p 1 has low abundance and is highly labile and was purified after expression as a recombinant protein in Escherichia coli. Pru p 3 is highly abundant in peach peel and was purified by conventional methods. The identities of the proteins were confirmed by sequence analysis and their masses determined by MS analysis. The purified proteins reacted with antisera against related allergens from other species: Pru p 1 with antiserum to Bet v 1 and Pru p 3 with antiserum to Mal d 3 (from apple). The presence of secondary and tertiary structure was demonstrated by circular dichroism (CD) and high field NMR spectroscopy. CD spectroscopy also showed that the two proteins differed in their stability at pH 3 and in their ability to refold after heating to 95 degrees C. Thus, Pru p 1 was unfolded at pH 3 even at 25 degrees C but was able to refold after heating to 95 degrees C at pH 7.5. In contrast, Pru p 3 was unable to refold after heating under neutral conditions but readily refolded after heating at pH 3.

KeywordsFood Science & Technology
Year of Publication2008
JournalMolecular Nutrition & Food Research
Journal citation52, pp. S220-S229
Digital Object Identifier (DOI)doi:10.1002/mnfr.200700274
PubMed ID18384093
Open accessPublished as non-open access
FunderBiotechnology and Biological Sciences Research Council
European Union - EU
Funder project or codeCentre for Crop Genetic Improvement (CGI)
ISSN16134125
PublisherWiley
Grant IDFOOD-CT-2005-514000

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