The human protein atlas blog


Proteomic analysis of cell cycle progression reveal mitotic substages

2017-11-02
Affinity proteomics Cell Atlas Cell cycle Immunofluorescence


Examples of spindle phenotypes (Bipole, Collapsed, Multipole and Monopole)

A key feature and a critical first step in understanding cell division and proliferation lies in characterizing the temporal regulation of protein abundance. A collaborative publication "Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS" was recently published in eLife.

The Cell atlas team from Sweden joined forces with Dr Tony Ly and Professor Angus Lamond from the University of Dundee, to perform a proteome-wide analysis of changes in protein abundance and phosphorylation across the cell cycle. This study reports the development of a novel technical approach, termed "PRoteomic analysis of Intracellular iMMUnolabelled cell Subsets" (PRIMMUS), that improves signal to noise in quantitative proteomics analyses by combining the separation of fixed and antibody stained cells by FACS with MS analysis of protein expression.

Using PRIMMUS enabled here the first detailed proteomic analysis of protein abundance variation in mitosis asynchronously growing cells, including the proteomic analysis of distinct mitotic substages. This identified a set of 136 proteins with significant abundance variation between different mitotic subphases. In addition, the study identified 115 phosphorylation sites increasing specifically during G2, prior to mitotic entry. All of the resulting MS data are available online, via the interactive EPD database at www.peptracker.com/epd Read the full article here.

References


Ly et al. Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS; doi: 10.7554/eLife.27574

Diana Mahdessian



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