Centrosome isolation and analysis by mass spectrometry-based proteomics

Lis Jakobsen, Jacob Morville Schrøder, Katja M Larsen, Emma Lundberg, Jens S. Andersen

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Abstrakt

Centrioles are microtubule-based scaffolds that are essential for the formation of centrosomes, cilia, and flagella with important functions throughout the cell cycle, in physiology and during development. The ability to purify centriole-containing organelles on a large scale, combined with advances in protein identification using mass spectrometry-based proteomics, have revealed multiple centriole-associated proteins that are conserved during evolution in eukaryotes. Despite these advances, the molecular basis for the plethora of processes coordinated by cilia and centrosomes is not fully understood. Considering the complexity and dynamics of centriole-related proteomes and the first-pass analyses reported so far, it is likely that further insight might come from more thorough proteome analyses under various cellular and physiological conditions. To this end, we here describe methods to isolate centrosomes from human cells and strategies to selectively identify and study the properties of the associated proteins using quantitative mass spectrometry-based proteomics.
OriginalsprogEngelsk
TitelMethods in Enzymology, Cilia, part B
RedaktørerWallace E. Marshall
Antal sider23
ForlagElsevier
Publikationsdato2013
Sider371-393
ISBN (Trykt)9780123979445
DOI
StatusUdgivet - 2013
NavnMethods in Enzymology
Vol/bind525
ISSN0076-6879

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