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

Sonja Gaier; Justin Marsh; Christina Oberhuber; Neil M. Rigby; Alison Lovegrove; Stefano Alessandri; Peter Briza; Christian Radauer; Laurian Zuidmeer; Ronald van Ree; Wolfgang Hemmer; Ana I. Sancho; Clare Mills; Karin Hoffmann-Sommergruber; Peter R. Shewry

doi:https://doi.org/10.1002/mnfr.200700274

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

Sonja Gaier, Justin Marsh, Christina Oberhuber, Neil M. Rigby, Alison Lovegrove, Stefano Alessandri, Peter Briza, Christian Radauer, Laurian Zuidmeer, Ronald van Ree, Wolfgang Hemmer, Ana I. Sancho, Clare Mills, Karin Hoffmann-Sommergruber, Peter R. Shewry

Research output: Contribution to journal › Article › Academic › peer-review

91 Citations (Scopus)

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

Original language	English
Pages (from-to)	S220-S229
Journal	Molecular nutrition & food research
Volume	52
Issue number	Suppl. 2
DOIs	https://doi.org/10.1002/mnfr.200700274
Publication status	Published - 2008

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https://doi.org/10.1002/mnfr.200700274

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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., & Shewry, P. R. (2008). Purification and structural stability of the peach allergens Pru p 1 and Pru p 3. Molecular nutrition & food research, 52(Suppl. 2), S220-S229. https://doi.org/10.1002/mnfr.200700274

@article{b44fce6078084ffea862b0f5d8001936,

title = "Purification and structural stability of the peach allergens Pru p 1 and Pru p 3",

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",

author = "Sonja Gaier and Justin Marsh and Christina Oberhuber and Rigby, {Neil M.} and Alison Lovegrove and Stefano Alessandri and Peter Briza and Christian Radauer and Laurian Zuidmeer and {van Ree}, Ronald and Wolfgang Hemmer and Sancho, {Ana I.} and Clare Mills and Karin Hoffmann-Sommergruber and Shewry, {Peter R.}",

year = "2008",

doi = "https://doi.org/10.1002/mnfr.200700274",

language = "English",

volume = "52",

pages = "S220--S229",

journal = "Molecular nutrition & food research",

issn = "1613-4125",

publisher = "Wiley-VCH Verlag",

number = "Suppl. 2",

}

Gaier, S, Marsh, J, Oberhuber, C, Rigby, NM, Lovegrove, A, Alessandri, S, Briza, P, Radauer, C, Zuidmeer, L , van Ree, R, Hemmer, W, Sancho, AI, Mills, C, Hoffmann-Sommergruber, K & Shewry, PR 2008, 'Purification and structural stability of the peach allergens Pru p 1 and Pru p 3', Molecular nutrition & food research, vol. 52, no. Suppl. 2, pp. S220-S229. https://doi.org/10.1002/mnfr.200700274

TY - JOUR

T1 - Purification and structural stability of the peach allergens Pru p 1 and Pru p 3

AU - Gaier, Sonja

AU - Marsh, Justin

AU - Oberhuber, Christina

AU - Rigby, Neil M.

AU - Lovegrove, Alison

AU - Alessandri, Stefano

AU - Briza, Peter

AU - Radauer, Christian

AU - Zuidmeer, Laurian

AU - van Ree, Ronald

AU - Hemmer, Wolfgang

AU - Sancho, Ana I.

AU - Mills, Clare

AU - Hoffmann-Sommergruber, Karin

AU - Shewry, Peter R.

PY - 2008

Y1 - 2008

N2 - 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

AB - 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

U2 - https://doi.org/10.1002/mnfr.200700274

DO - https://doi.org/10.1002/mnfr.200700274

M3 - Article

C2 - 18384093

SN - 1613-4125

VL - 52

SP - S220-S229

JO - Molecular nutrition & food research

JF - Molecular nutrition & food research

IS - Suppl. 2

ER -