title: Protein extraction from plant tissues for 2DE and its application in proteomic analysis.
abstract: Plant tissues contain large amounts of secondary compounds that significantly interfere with protein extraction and 2DE analysis. Thus, sample preparation is a crucial step prior to 2DE in plant proteomics. This tutorial highlights the guidelines that need to be followed to perform an adequate total protein extraction before 2DE in plant proteomics. We briefly describe the history, development, and feature of major sample preparation methods for the 2DE analysis of plant tissues, that is, trichloroacetic acid/acetone precipitation and phenol extraction. We introduce the interfering compounds in plant tissues and the general guidelines for tissue disruption, protein precipitation and resolubilization. We describe in details the advantages, limitations, and application of the trichloroacetic acid/acetone precipitation and phenol extraction methods to enable the readers to select the appropriate method for a specific species, tissue, or cell type. The current applications of the sample preparation methods in plant proteomics in the literature are analyzed. A comparative proteomic analysis between male and female plants of Pistacia chinensis is used as an example to represent the sample preparation methodology in 2DE-based proteomics. Finally, the current limitations and future development of these sample preparation methods are discussed. This Tutorial is part of the International Proteomics Tutorial Programme (IPTP17).
authors.affiliation: State Key Laboratory of Wheat & Maize Crop Science in Henan Province, Synergetic Innovation Center of Henan Grain Crops, College of Life Science, Henan Agricultural University, Zhengzhou, China.
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Wu
authors.forename: Fangping
authors.initials: F
authors.lastname: Gong
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D000096:Acetone / Q000737:chemistry; D011232:Chemical Precipitation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D000432:Methanol / Q000737:chemistry; D010636:Phenols / Q000737:chemistry; D027927:Pistacia / Q000737:chemistry*; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification*; D010944:Plants / Q000737:chemistry*; D020543:Proteome / Q000032:analysis / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*
chemical_list: D010636:Phenols; D010940:Plant Proteins; D020543:Proteome; D000096:Acetone; D000432:Methanol
keywords: 2DE; Plant proteomics; Sample preparation; Tutorial