Authors: Eubel, Holger¹; Lee, Chun Pong; Kuo, John²; Meyer, Etienne H.¹; Taylor, Nicolas L.¹; Millar, A. Harvey

Source: The Plant Journal, Volume 52, Number 3, November 2007 , pp. 583-594(12)

Publisher: Wiley-Blackwell

Abstract:

Summary

Sample purity is the key for a successful in-depth analysis of any given subcellular proteome. The suitability of free-flow electrophoresis to assist conventional, centrifugation-based techniques in the preparation of plant mitochondria from green and non-green tissue was assessed by various means, including functional assays, immunoblots, electron microscopy and differential gel electrophoresis. Results indicated a significant increase in purity of the mitochondrial samples, highlighted specific contaminants previously reported as mitochondrial proteins, and also pointed to new means for separating plastids and peroxisomes from mitochondria in plant organellar extracts by exploiting differences in surface charge. This approach has the potential to allow a deeper and more comprehensive investigation of the Arabidopsis organellar proteomes, by providing a second dimension of separation based on surface charge in addition to conventional centrifugation purification protocols relying on size and density.

Keywords: plant mitochondria; proteomics; sample purity; organellar contamination; free-flow electrophoresis; centrifugation

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1365-313X.2007.03253.x

Affiliations: 1: ARC Centre of Excellence in Plant Energy Biology, The University of Western Australia, 35 Stirling Highway, M316, Crawley, WA 6009, Australia, 2: School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia, Stirling Highway, M310, Crawley, WA 6009, Australia, and

Publication date: 2007-11-01

Related content

• In this: publication
• By this: publisher
• In this Subject: Botany
• By this author: Eubel, Holger ;  Lee, Chun Pong ;  Kuo, John ;  Meyer, Etienne H. ;  Taylor, Nicolas L. ;  Millar, A. Harvey

Website © Publishing Technology. Article copyright remains with the publisher, society or author(s) as specified within the article.

• Terms and conditions
• Privacy policy
• Information for advertisers