Beyond the Western front: targeted proteomics and organelle abundance profiling.

Harriet T Parsons,Joshua L Heazlewood

doi:10.3389/fpls.2015.00301

Abstract

The application of westerns or immunoblotting techniques for assessing the composition, dynamics, and purity of protein extracts from plant material has become common practice. While the approach is reproducible, can be readily applied and is generally considered robust, the field of plant science suffers from a lack of antibody variety against plant proteins. The development of approaches that employ mass spectrometry to enable both relative and absolute quantification of many hundreds of proteins in a single sample from a single analysis provides a mechanism to overcome the expensive impediment in having to develop antibodies in plant science. We consider it an opportune moment to consider and better develop the adoption of multiple reaction monitoring (MRM)-based analyses in plant biochemistry.

Highlights

The application of westerns or immunoblotting techniques for assessing the composition, dynamics, and purity of protein extracts from plant material has become common practice
For compartments that were less enriched than discrete organelles like the plastid or mitochondrion, assessment of contamination levels became more pressing and researchers turned toward immunoblotting, as well as enzyme assays and microscopy (Norman et al, 1986; Hahn et al, 1987; Meyer et al, 1988)
The advent of modern mass spectrometry and proteomics meant that could the main biochemical reactions or constituents of a compartment be investigated but many potentially functionally associated proteins could be identified, making organelle proteomics a valuable tool for Targeted plant organelle proteomics reducing the complexity of the eukaryotic cell

Summary

Introduction

The application of westerns or immunoblotting techniques for assessing the composition, dynamics, and purity of protein extracts from plant material has become common practice. The development of approaches that employ mass spectrometry to enable both relative and absolute quantification of many hundreds of proteins in a single sample from a single analysis provides a mechanism to overcome the expensive impediment in having to develop antibodies in plant science.

Objectives

Results

Conclusion