Repository hosted by TU Delft Library

Home · Contact · About · Disclaimer ·
 

Isolation and characterization of patatin isoforms

Publication files not online:

Author: Pots, A.M. · Gruppen, H. · Hessing, M. · Boekel, M.A.J.S. van · Voragen, A.G.J.
Type:article
Date:1999
Source:Journal of Agricultural and Food Chemistry, 11, 47, 4587-4592
Identifier: 235198
doi: doi:10.1021/jf981180n
Keywords: Characterization · Isolation · Patatin isoforms · patatin · protein · unclassified drug · vegetable protein · article · capillary electrophoresis · chemistry · circular dichroism · conformation · fractionation · high performance liquid chromatography · isoelectric focusing · isolation and purification · mass spectrometry · multigene family · physical chemistry · polyacrylamide gel electrophoresis · protein analysis · protein conformation · protein folding · protein isolation · protein stability · protein structure · structure analysis · temperature · Carboxylic Ester Hydrolases · Chromatography, High Pressure Liquid · Circular Dichroism · Plant Proteins · Protein Conformation · Protein Folding · Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization · Temperature

Abstract

Patatin has, so far, been considered a homogeneous group of proteins. A comparison of the isoforms in terms of structural properties or stability has not been reported. A method to obtain various isoform fractions as well as a comparison of the physicochemical properties of these pools is presented. Patatin could be separated in four isoform pools, denoted A, B, C, and D, representing 62%, 26%, 5%, and 7% of the total amount of patatin, respectively. These isoforms differed in surface charge, resulting in a different behavior on anion exchange chromatography, isoelectric focusing, native polyacrylamide gel, and capillary electrophoresis. All isoforms of the patatin family contained proteins with two molecular masses of approximately 40.3 and 41.6 kDa, respectively. The size of this difference in the molar mass (1300 Da) is on the order of one carbohydrate moiety. Despite the biochemical differences given above, no variations in the structural properties nor in the thermal conformational stability could be observed using far-ultraviolet circular dichroism, infrared, and fluorescence spectroscopy.