title: [Extraction of cytoskeletons and associated proteins using subcellular proteome fractionation technique].
abstract: OBJECTIVE
  To develop a method for extracting cytoskeletons and cytoskeleton-associated proteins for proteomic analysis.
  
  
  METHODS
  A subcellular sequential proteome extraction method was exploited. The extraction procedure was optimized and controlled according to observed cell morphology changes and one- and two-dimensional electrophoresis images. The extraction efficiency and selectivity were evaluated by Western blotting and mass spectrometry.
  
  
  RESULTS
  Four extracted fractions clearly displayed distinct patterns. Western blotting detected the fraction-marker proteins FAK, integrin-β1, histone H1 and cytokeratin 19 only in their expected fractions. About 90% of the protein spots in the cytoskeleton fraction were identified by mass spectrometry as cytoskeleton and/or its associated proteins.
  
  
  CONCLUSIONS
  The subcellular proteome sequential fractionation method facilitates the detection of proteins of low abundance and shows a high reproducibility and selectivity, and thus can serve as an ideal pre-fractionation method prior to two-dimensional electrophoresis.
authors.affiliation: Southern Medical University, Guangzhou, China. jean.cz@qq.com
authors.forename: Zhen
authors.initials: Z
authors.lastname: Cai
authors.forename: Shilong
authors.initials: S
authors.lastname: Xiong
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Huicai
authors.initials: H
authors.lastname: Ji
authors.forename: Chunyan
authors.initials: C
authors.lastname: Xinag
authors.forename: Lei
authors.initials: L
authors.lastname: Zheng
mesh_terms: D003599:Cytoskeleton / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D013347:Subcellular Fractions
chemical_list: D020543:Proteome