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Purification and structural characterisation of lipid transfer protein from red wine and grapes
Authors:Nadine Jaeckels  Stefan Tenzer  Susanne Rosfa  Hansjoerg Schild  Heinz Decker  Petra Wigand
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
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.
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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