Purification of Target Proteins from Native Tissues: CCT Complex from Bovine Testes and PP2Ac from Porcine Brains

Methods Mol Biol. 2018:1788:73-88. doi: 10.1007/7651_2017_89.

Abstract

Proteomics is a powerful approach for systematic identification and quantification of the entire proteome of a biological system (cell, tissue, organ, biological fluid, or organism) at specific time points ( http://www.nature.com ). Extracting and purifying target proteins from native tissues are essential steps for many aspects of proteomic studies. In this chapter, we will introduce the experimental procedures to obtain soluble proteins from two different tissues: (1) the CCT (cpn-containing TCP-1) complex from bovine testes and (2) the protein phosphatase 2A (PP2A) catalytic subunit (PP2Ac or C) from porcine brains. With these two examples, we would like to provide some general guidelines for researchers on how to extract and purify target proteins from specific tissues and extend these approaches to other proteins of interest.

Keywords: Bovine testes; CCT; FPLC; PP2A; Porcine brains; Protein purification; Proteomic.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Brain Chemistry*
  • Catalytic Domain
  • Cattle
  • Chaperonin Containing TCP-1 / chemistry
  • Chaperonin Containing TCP-1 / isolation & purification*
  • Chromatography, Ion Exchange / methods
  • Electrophoresis, Polyacrylamide Gel / methods
  • Male
  • Protein Phosphatase 2 / chemistry
  • Protein Phosphatase 2 / isolation & purification*
  • Protein Subunits / chemistry
  • Protein Subunits / isolation & purification
  • Proteomics / methods*
  • Solubility
  • Swine
  • Testis / chemistry*

Substances

  • Protein Subunits
  • Protein Phosphatase 2
  • Chaperonin Containing TCP-1