Sequential solubilization of proteins precipitated with trichloroacetic acid in acetone from cultured Catharanthus roseus cells yields 52% more spots after two-dimensional electrophoresis

Proteomics
D I JacobsR Verpoorte

Abstract

Sample preparation is still the most critical step in two-dimensional gel electrophoresis (2-DE), and needs to be optimized for each type of sample. To analyze the proteome of the medicinal plant Catharanthus roseus, we developed and evaluated a sequential solubilization procedure for the solubilization of proteins after precipitation in trichloroacetic acid and acetone. The procedure includes solubilization with a conventional urea buffer followed by a stronger solubilizing buffer containing thiourea. The sequential solubilization of the precipitated proteins results in very different spot patterns following 2-DE. The number of protein spots which could be detected in both samples of the sequential solubilization was only about 10% of the total number of spots. Compared to solubilization in a single step, the total number of spots that could be detected in the sequential solubilization procedure was increased by 52%. The method described is simple and is applicable to different types of plant tissue.

Citations

Jul 9, 2002·Phytochemical Analysis : PCA
Feb 1, 2005·Planta·Denise I JacobsRobert Verpoorte
Aug 1, 2010·Mycotoxin Research·Muhammad RizwanEbrahim Razzazi-Fazeli
Aug 31, 2010·Transactions of the Royal Society of Tropical Medicine and Hygiene·E-G LeeY Kong
Dec 17, 2009·Phytochemistry·B F QuirinoR H Krüger
Aug 19, 2004·The Plant Journal : for Cell and Molecular Biology·Jocelyn K C RoseRamu Subramanian Saravanan
Nov 5, 2004·Proteomics·Manuela BassoMauro Fasano
Apr 24, 2009·Protein Science : a Publication of the Protein Society·Dakshinamurthy RajalingamThallapuranam Krishnaswamy S Kumar
Jun 16, 2005·Mass Spectrometry Reviews·Ganesh Kumar Agrawal, Randeep Rakwal
Oct 17, 2009·Proteomics·Ingrid MillerElisabetta Gianazza
Jul 28, 2004·Phytochemistry·Eric SarnighausenRalf Reski
Sep 9, 2006·International Journal for Parasitology·Patrick de VenevellesMarie Labbé
Jan 17, 2017·Journal of Proteome Research·Melda Z GürayAlan A Doucette
Jul 11, 2009·Molecular Plant-microbe Interactions : MPMI·Eugenia Morán-DiezEnrique Monte
Oct 1, 2008·Journal of Phycology·Loretto ContrerasJuan A Correa

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