Proteomics of organelles and large cellular structures

Abstract

The mass-spectrometry-based identification of proteins has created opportunities for the study of organelles, transport intermediates and large subcellular structures. Traditional cell-biology techniques are used to enrich these structures for proteomics analyses, and such analyses provide insights into the biology and functions of these structures. Here, we review the state-of-the-art proteomics techniques for the analysis of subcellular structures and discuss the biological insights that have been derived from such studies.

Quantitative Organelle Proteomics of MCF-7 Breast Cancer Cells Reveals Multiple Subcellular Locations for Proteins in Cellular Functional Processes

“…[24][25][26][27][28] The most common approach has been purification of individual organelles followed by exhaustive determination of the protein content. The main disadvantages of this approach are that the degree of purification/ contamination of the organelle is difficult to ascertain conclusively for lower abundance proteins, that the protein content may be altered by the purification process and that the approach is not very suitable for dynamic studies of protein subcellular location.…”

Subcellular proteomics of mice gastrocnemius and soleus muscles

“…In a recent proteomics study on murine gastrocnemius and soleus muscle extracts, performed by Gelfi and coworkers [9], more than 800 spots on each 2-DE were detected by silver staining, leading to the identification of 85 different proteins belonging to the most abundant structural and metabolic protein classes. Despite the large number of visualized spots by 2-DE, proteins with lower relative abundances, usually involved in protein biosynthesis and cell stress response, are probably masked by structural or metabolic proteins [18,19]. To counteract this, Jarrold and coworkers [14] performed the depletion of abundant muscle proteins by a high pH treatment followed by 2-DE analysis, resulting in the elimination of the major contractile proteins and in the detection of nine minor proteins, mainly belonging to mitochondria, for the first time.…”

Proteomics in human cancer research

“…This study combines large-scale proteomic analysis with classical cell biology methods, and this is supplemented by novel centrifugation and affinity-mediated techniques [105,106]. Proteomic analyses combined with protein localization and protein-knock-down techniques could identify the phenotype, which is produced by disruption of the protein, and elucidate the function of organelle and the individual protein in disease.…”