Purification and structural characterisation of lipid transfer protein from red wine and grapes |
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Authors: | Nadine Jaeckels Stefan Tenzer Susanne Rosfa Hansjoerg Schild Heinz Decker Petra Wigand |
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Affiliation: | 1. Institute for Molecular Biophysics, Johannes Gutenberg-University Mainz, Jakob Welder-Weg 26, 55128 Mainz, Germany;2. Institute for Immunology, University Medical Center of the Johannes Gutenberg-University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany |
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Abstract: | Lipid transfer proteins (LTP) play a major role in plant defence and are of particular interest due to their known ability to cause allergic reactions. These proteins are expressed in grapes and also remain detectable after vinification, especially in red wine. However, it remains unknown whether the protein undergoes any changes during the vinification process. Here, we present a purification method for LTPs from Dornfelder grapes and wine. By liquid-chromatography–mass spectroscopy (LC–MS/MS) we identified LTPs from two different species (Vitis vinifera and Vitis aestivalis). Additionally, the purified LTPs were characterised using spectrometric methods, confirming their high purity and structural stability during vinification. We conclude that LTPs are resistant to the alcohol content (13.5 vol%), acidic milieu of wine and other ingredients present during the vinification process, indicating that the allergenic potential of grape LTP is not diminished by the vinification process. |
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Keywords: | CD circular dichroism DTT dithiothreitol EDTA ethylenediaminetetraacetate ESI electrospray ionisation FPLC fast protein liquid chromatography IEP isoelectric point LC&ndash MS/MS liquid-chromatography&ndash mass spectroscopy LTP lipid transfer protein NaCl sodium chloride NaN3 sodium azide PDB protein data base PVP polyvinylpyrrolidone SDS sodium dodecyl sulfate SDS&ndash PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis TOF time-of-flight UPLC ultra performance liquid chromatography |
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