Evaluation of chloroform/methanol extraction to facilitate the study of membrane proteins of non-model plants

• Authors : Vertommen, A.; Panis, B.; Swennen, R.; Carpentier, S.


• Document type : Journal article


• Year of publication : 2010


• Journal title : Planta


• Volume (number) : 231 (5)




• Pages : 1113-1125


• Peer-reviewed : Yes


• ISSN : 0032-0935; 1432-2048


• Language(s) : English


• Abstract : Membrane proteins are of great interest to plant physiologists because of their important function in many physiological processes. However, their study is hampered by their low abundance and poor solubility in aqueous buffers. Proteomics studies of non-model plants are generally restricted to gel-based methods. Unfortunately, all gel-based techniques for membrane proteomics lack resolving power. Therefore, a very stringent enrichment method is needed before protein separation. In this study, protein extraction in a mixture of chloroform and methanol in combination with gel electrophoresis is evaluated as a method to study membrane proteins in non-model plants. Benefits as well as disadvantages of the method are discussed. To demonstrate the pitfalls of working with non-model plants and to give a proof of principle, the method was first applied to whole leaves of the model plant Arabidopsis.
  
  [Membrane proteins are of great interest to plant physiologists because of their important function in many physiological processes. However, their study is hampered by their low abundance and poor solubility in aqueous buffers. Proteomics studies of non-model plants are generally restricted to gel-based methods. Unfortunately, all gel-based techniques for membrane proteomics lack resolving power. Therefore, a very stringent enrichment method is needed before protein separation. In this study, protein extraction in a mixture of chloroform and methanol in combination with gel electrophoresis is evaluated as a method to study membrane proteins in non-model plants. Benefits as well as disadvantages of the method are discussed. To demonstrate the pitfalls of working with non-model plants and to give a proof of principle, the method was first applied to whole leaves of the model plant Arabidopsis.]


• Keywords : PLANT PHYSIOLOGY; LEAVES; ELECTROPHORESIS; PROTEINS; MERISTEMS; EXTRACTION; SIMULATION MODELS; JELLIFICATION; ARABIDOPSIS THALIANA; METHANOL


• Open access : Yes


• Document on publisher's site : View article on publisher's site


• Musalit document ID : IN100259

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