Chloroform-assisted phenol extraction improving proteome profiling of maize embryos through selective depletion of high-abundance storage proteins.
The presence of abundant storage proteins in plant embryos greatly impedes seed proteomics analysis. Vicilin (or globulin-1) is the most abundant storage protein in maize embryo. There is a need to deplete the vicilins from maize embryo extracts for enhanced proteomics analysis. We here reported a chloroform-assisted phenol extraction (CAPE) method for vicilin depletion. By CAPE, maize embryo proteins were first extracted in an aqueous buffer, denatured by chloroform and then subjected to phenol extraction. We found that CAPE can effectively deplete the vicilins from maize embryo extract, allowing the detection of low-abundance proteins that were masked by vicilins in 2-DE gel. The novelty of CAPE is that it selectively depletes abundant storage proteins from embryo extracts of both monocot (maize) and dicot (soybean and pea) seeds, whereas other embryo proteins were not depleted. CAPE can significantly improve proteome profiling of embryos and extends the application of chloroform and phenol extraction in plant proteomics. In addition, the rationale behind CAPE depletion of abundant storage proteins was explored.
Collaborative Innovation Center of Henan Grain Crops, College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Erhui
E
Xiong
Collaborative Innovation Center of Henan Grain Crops, College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Xiaolin
X
Wu
Collaborative Innovation Center of Henan Grain Crops, College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Le
L
Yang
Collaborative Innovation Center of Henan Grain Crops, College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Fangping
F
Gong
Collaborative Innovation Center of Henan Grain Crops, College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Fuju
F
Tai
Collaborative Innovation Center of Henan Grain Crops, College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Wei
W
Wang
D000595:Amino Acid Sequence; D005591:Chemical Fractionation / Q000379:methods*; D002725:Chloroform / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D008969:Molecular Sequence Data; D019800:Phenol / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D055314:Seed Storage Proteins / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry*; D013025:Glycine max / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D003313:Zea mays / Q000737:chemistry*
D055314:Seed Storage Proteins; D019800:Phenol; D002725:Chloroform; C015575:vicilin protein, plant