A protein extraction method compatible with proteomic analysis for the euhalophyte Salicornia europaea.
Protein extraction from plants like the halophyte Salicornia europaea has been problematic using standard protocols due to high concentrations of salt ions in their cells. We have developed an improved method for protein extraction from S. europaea, which allowed us to remove interfering compounds and salt ions by including the chemicals borax, polyvinylpolypyrrolidone, and phenol. The comparative study of this method with several other protocols using NaCl-treated S. europaea shoots demonstrated that this method gave the best distinction of proteins on 2-DE gels. This protocol had a wide range of applications as high yields and good distinction of 1-DE gels for proteins isolated from twelve other plants were rendered. In addition, we reported results of 2-DE using the recalcitrant tissue of the S. europaea roots. We also demonstrated that this protocol is compatible with proteomic analysis as eight specific proteins generated by this method have been identified by MS. In conclusion, our newly developed protein extraction protocol is expected to have excellent applications in proteomic studies of halophytes.
Wang
Xuchu
X
Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences Beijing, P.R. China.
Li
Xiaofang
X
Deng
Xin
X
Han
Heping
H
Shi
Wuliang
W
Li
Yinxin
Y
D001881:Borates / Q000737:chemistry; D002457:Cell Extracts / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D019663:Chenopodiaceae / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D013058:Mass Spectrometry / Q000379:methods*; D008623:Mercaptoethanol / Q000737:chemistry; D017830:Octoxynol / Q000737:chemistry; D019800:Phenol / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011205:Povidone / Q000031:analogs & derivatives / Q000737:chemistry; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods; D012965:Sodium Chloride / Q000737:chemistry / Q000302:isolation & purification; D012997:Solvents / Q000737:chemistry
D001881:Borates; D002457:Cell Extracts; D010940:Plant Proteins; D020543:Proteome; D012997:Solvents; C018851:borax; C077842:polyvinylpolypyrrolidone; D019800:Phenol; D012965:Sodium Chloride; D008623:Mercaptoethanol; D017830:Octoxynol; D011205:Povidone