Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Issue 1 (3rd May 2016)

• Record Type: Journal Article
• Authors: LaMontagne, Erica D.; Collins, Carina A.; Peck, Scott C.; Heese, Antje
• Editors: Birchler, James; Ecker, Joseph; Martin, Cathie R.; Stitt, Mark; Zhou, Jian-Min
• Other Names: Stacey Gary editorInChief.
• Is Part Of: Current protocols in plant biology. Volume 1:Issue 1(2016)
• Page Start: 217
• Page End: 234
• Publication Date: 2016-05-03
• Subjects: Arabidopsis thaliana -- cellular fractionation -- differential centrifugation -- microsomes -- solubilization of microsomal proteins; Botany -- Laboratory manuals; Plants; Botany; Botany; Laboratory Manual; Periodical; Laboratory manuals; 580.28
• DOI: 10.1002/cppb.20020
• Languages: English
• ISSNs: 2379-8068

Abstract

Abstract: Cellular membranes define the boundaries between organelles and the cytosol or the extracellular environment, thus providing functional separation between subcellular compartments. In addition, membranes assist in a diverse range of cellular functions, including serving as signaling platforms, mediating transport of molecules, and facilitating trafficking of cargo between cellular compartments. Because membrane functionality is largely defined by protein composition, exploring the roles of membrane proteins is of interest to many researchers. This article focuses on the subcellular fractionation of microsomes, which are membrane‐derived vesicles formed during cell lysis. In plants, microsomes mainly consist of the plasma membrane and membranes derived from the endoplasmic reticulum, Golgi apparatus, trans‐Golgi network, and tonoplast. The article describes the different steps involved in enriching for and solubilizing microsomal membrane proteins from Arabidopsis thaliana seedlings and cultured cells by differential centrifugation. Solubilized microsomal proteins can be used for subsequent immunoblot analysis, co‐immunoprecipitation, or proteomic studies. © 2016 by John Wiley & Sons, Inc.