Isolation and characterization of patatin isoforms.
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.
Pots
A M
AM
Centre for Protein Technology TNO-WAU, Wageningen, The Netherlands.
Gruppen
H
H
Hessing
M
M
van Boekel
M A
MA
Voragen
A G
AG
D002265:Carboxylic Ester Hydrolases; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D011487:Protein Conformation; D017510:Protein Folding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013696:Temperature
D010940:Plant Proteins; C043222:patatin protein, Solanum tuberosum; D002265:Carboxylic Ester Hydrolases