RRC ID 11877
Author Nouri MZ, Komatsu S.
Title Comparative analysis of soybean plasma membrane proteins under osmotic stress using gel-based and LC MS/MS-based proteomics approaches.
Journal Proteomics
Abstract To study the soybean plasma membrane proteome under osmotic stress, two methods were used: a gel-based and a LC MS/MS-based proteomics method. Two-day-old seedlings were subjected to 10% PEG for 2 days. Plasma membranes were purified from seedlings using a two-phase partitioning method and their purity was verified by measuring ATPase activity. Using the gel-based proteomics, four and eight protein spots were identified as up- and downregulated, respectively, whereas in the nanoLC MS/MS approach, 11 and 75 proteins were identified as up- and downregulated, respectively, under PEG treatment. Out of osmotic stress responsive proteins, most of the transporter proteins and all proteins with high number of transmembrane helices as well as low-abundance proteins could be identified by the LC MS/MS-based method. Three homologues of plasma membrane H(+)-ATPase, which are transporter proteins involved in ion efflux, were upregulated under osmotic stress. Gene expression of this protein was increased after 12 h of stress exposure. Among the identified proteins, seven proteins were mutual in two proteomics techniques, in which calnexin was the highly upregulated protein. Accumulation of calnexin in plasma membrane was confirmed by immunoblot analysis. These results suggest that under hyperosmotic conditions, calnexin accumulates in the plasma membrane and ion efflux accelerates by upregulation of plasma membrane H(+)-ATPase protein.
Volume 10(10)
Pages 1930-45
Published 2010-5
DOI 10.1002/pmic.200900632
PMID 20209511
MeSH Chromatography, Liquid Electrophoresis, Gel, Two-Dimensional Gene Expression Regulation, Plant Hypocotyl / growth & development Membrane Proteins / analysis* Membrane Proteins / genetics Osmosis* Plant Roots / growth & development Proteomics Soybean Proteins / analysis* Soybean Proteins / genetics Soybeans / chemistry* Soybeans / genetics Soybeans / growth & development Stress, Physiological* Tandem Mass Spectrometry
IF 3.532
Times Cited 55
WOS Category BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Lotus / Glycine GMFLcDNA