Proteomic profiling of centrosomes across multiple mammalian cell and tissue types by an affinity capture method
The lab of Fanni Gergely has developed a new affinity-based method that allows rapid isolation of centrosomes for proteomic profiling.
Centrosomes are best known for nucleating and organising the microtubule cytoskeleton in proliferating cells. They play fundamental roles in a variety of cellular pathways ranging from mitotic spindle formation, trafficking and signaling to cilia assembly, cellular organization and polarity. Accordingly, congenital mutations in centrosomal genes lead to a variety of developmental disorders and pathologies.
Being small membraneless organelles, centrosomes are built of hundreds of different proteins; although the centrosome proteome was successfully deciphered in a couple of cell lines, we still lack the tools for systematic analysis of centrosome composition during dynamic processes or across cell types. Moreover, we do not know if centrosomes contain cell type- or tissue-specific components, which could have important implications to our understanding of centrosome function.
Using a biotinylated peptide derived from the centrosomal protein CCDC61 the Gergely lab and their collaborators, Drs Takashi Ochi and Mark van Breugel, have developed a rapid single-step technique to purify centrosomes for mass-spectrometry analysis. This new method, which they named CAPture, provides much greater specificity and yield than current alternative approaches.
"With CAPture we were finally able to characterise centrosome proteomes across different cell types. Not only did we unveil previously unseen heterogeneity in centrosome composition, using this information we could also identify new hierarchical interactions within the centrosome"
The authors believe that CAPture combined with mass-spectrometry constitutes a transformative tool to explore dynamic changes in centrosome proteomes in health and disease.
Fanni Gergely
10th October 2023
