Abstract
Protein phosphorylation is one of the most common post-translational modifications in eukaryotic cells and plays a critical role in a vast array of cellular processes. Efficient methods of protein extraction and phosphopeptide purification are required to ensure the detection of high quality of proteins. In our hands, phenol extraction of proteins and TiO2 chromatography enrich phosphorylated peptides more efficiently than other methods in the moss Physcomitrlla patens (P. patens).
Materials and Reagents
Equipment
Procedure
This procedure includes proteins extraction, protein digestion and phosphopeptide purification. The following should all be carried out at cold room temperature except for special instructions.
Recipes
Acknowledgments
This work was supported by grants from Project of Ministry Science & Technology of China (2013CB967300, 2007CB948201) to Prof. He, grant from National Natural Science Foundation of China (No. 31371243, 30970250), China Postdoctoral Science Foundation and State Education Ministry Scientific Research Foundation for the Returned Overseas Chinese Scholars to Dr. Wang.
References
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