Author's School

Arts & Sciences

Author's Department

Biology

Document Type

Article

Publication Date

10-12-2016

Originally Published In

Isolation of Plant Organelles and Structures, Volume 1511 of the series Methods in Molecular Biology pp 301-334. DOI: 10.1007/978-1-4939-6533-5_24

Abstract

The 26S proteasome is a highly dynamic, multisubunit, ATP-dependent protease that plays a central role in cellular housekeeping and many aspects of plant growth and development by degrading aberrant polypeptides and key cellular regulators that are first modified by ubiquitin. Although the 26S proteasome was originally enriched from plants over 30 years ago, only recently have significant advances been made in our ability to isolate and study the plant particle. Here, we describe two robust methods for purifying the 26S proteasome and its subcomplexes from Arabidopsis thaliana; one that involves conventional chromatography techniques to isolate the complex from wild-type plants, and another that employs the genetic replacement of individual subunits with epitope-tagged variants combined with affinity purification. In addition to these purification protocols, we describe methods commonly used to analyze the activity and composition of the complex.

Comments

Accepted manuscript version of chapter from Isolation of Plant Organelles and Structures, Volume 1511 of the series Methods in Molecular Biology pp 301-334. DOI: 10.1007/978-1-4939-6533-5_24

DOI

10.1007/978-1-4939-6533-5_24

Embargo Period

10-12-2017

Available for download on Thursday, October 12, 2017

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