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Issue 13 (2) 2014 pp. 181-190

Jerzy Dziuba, Iwona Szerszunowicz, Dorota Nałęcz, Marta Dziuba

Department of Food Biochemistry, University of Warmia and Mazury in Olsztyn, Poland

Proteomic analysis of albumin and globulin fractions of pea (Pisum sativum L.) seeds

Abstract

Background. Proteomic analysis is emerging as a highly useful tool in food research, including studies of food allergies. Two-dimensional gel electrophoresis involving isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis is the most effective method of separating hundreds or even thousands of proteins. In this study, albumin and globulin tractions of pea seeds cv. Ramrod were subjected to proteomic analysis. Selected potentially alergenic proteins were identified based on their molecular weights and isoelectric points.
Material and methods. Pea seeds (Pisum sativum L.) cv. Ramrod harvested over a period of two years (Plant Breeding Station in Piaski-Szelejewo) were used in the experiment. The isolated albumins, globulins and legumin and vicilin fractions of globulins were separated by two-dimensional gel electrophoresis. Proteomic images were analysed in the ImageMaster 2D Platinum program with the use of algorithms from the Melanie application. The relative content, isoelectric points and molecular weights were computed for
all identified proteins. Electrophoregrams were analysed by matching spot positions from three independent replications.
Results. The proteomes of albumins, globulins and legumin and vicilin fractions of globulins produced up to several hundred spots (proteins). Spots most characteristic of a given fraction were identified by computer analysis and spot matching. The albumin proteome accumulated spots of relatively high intensity over a broad range of pi values of -4.2-8.1 in 3 molecular weight (MW) ranges: I - high molecular-weight albumins with MW of -50-110 kDa, II - average molecular-weight albumins with MW of -20-35 kDa, and III - low molecular-weight albumins with MW of -13-17 kDa. 2D gel electrophoregrams revealed the presence of 81 characteristic spots, including 24 characteristic of legumin and 14 - of vicilin.
Conclusions. Two-dimensional gel electrophoresis proved to be a useful tool for identifying pea proteins. Patterns of spots with similar isoelectric points and different molecular weights or spots with different isoelectric points and similar molecular weights play an important role in proteome analysis. The regions characteristic of albumin, globulin and legumin and vicilin fractions of globulin with typical MW and pi values were identified as the results of performed 2D electrophoretic separations of pea proteins. 2D gel electrophoresis of albumins and the vicilin fraction of globulins revealed the presence of 4 and 2 spots, respectively, representing potentially allergenic proteins. They probably corresponded to vicilin fragments synthesized during post-translational modification of the analysed protein.

Keywords: pea seed proteins, 2D-PAGE, allergenic proteins
pub/.pdf Full text available in english in Adobe Acrobat format:
http://www.food.actapol.net/issue2/volume/7_2_2014.pdf

http://dx.doi.org/10.17306/J.AFS.2014.2.7

For citation:

MLA Dziuba, Jerzy, et al. "Proteomic analysis of albumin and globulin fractions of pea (Pisum sativum L.) seeds." Acta Sci.Pol. Technol. Aliment. 13.2 (2014): 181-190. http://dx.doi.org/10.17306/J.AFS.2014.2.7
APA Dziuba J., Szerszunowicz I., Nałęcz D., Dziuba M., (2014). Proteomic analysis of albumin and globulin fractions of pea (Pisum sativum L.) seeds. Acta Sci.Pol. Technol. Aliment. 13 (2), 181-190 http://dx.doi.org/10.17306/J.AFS.2014.2.7
ISO 690 DZIUBA, Jerzy, et al. Proteomic analysis of albumin and globulin fractions of pea (Pisum sativum L.) seeds. Acta Sci.Pol. Technol. Aliment., 2014, 13.2: 181-190. http://dx.doi.org/10.17306/J.AFS.2014.2.7