Grape and wine proteins: their fractionation by hydrophobic interaction chromatography and identification by chromatographic and proteomic analysis.
A method to fractionate grape and wine proteins by hydrophobic interaction chromatography (HIC) was developed. This method allowed the isolation of a thaumatin-like protein in a single step with high yield and >90% purity and has potential to purify several other proteins. In addition, by separating HIC fractions by reverse phase HPLC and by collecting the obtained peaks, the grape juice proteins were further separated, by SDS-PAGE, into 24 bands. The bands were subjected to nanoLC-MS/MS analysis, and the results were matched against a database and characterized as various Vitis vinifera proteins. Moreover, either directly or by homology searching, identity or function was attributed to all of the gel bands identified, which mainly consisted of grape chitinases and thaumatin-like proteins but also included vacuolar invertase, PR-4 type proteins, and a lipid transfer protein from grapes.
Marangon
Matteo
M
Dipartimento di Biotecnologie agrarie and Centro Interdipartimentale per la Ricerca in Viticoltura ed Enologia, Università di Padova, via dell'Università 16, I-35020 Legnaro, PD, Italy.
Van Sluyter
Steven C
SC
Haynes
Paul A
PA
Waters
Elizabeth J
EJ
D052179:Antigens, Plant / Q000032:analysis; D002352:Carrier Proteins / Q000032:analysis; D005591:Chemical Fractionation / Q000379:methods; D002688:Chitinases / Q000032:analysis; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D005638:Fruit / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D010940:Plant Proteins / Q000032:analysis* / Q000302:isolation & purification*; D027843:Vitis / Q000737:chemistry*; D014920:Wine / Q000032:analysis*; D043324:beta-Fructofuranosidase / Q000032:analysis
D052179:Antigens, Plant; D002352:Carrier Proteins; D010940:Plant Proteins; C511920:lipid transfer proteins, plant; C003427:thaumatin protein, plant; D002688:Chitinases; D043324:beta-Fructofuranosidase