Proteomic profiling of centrosomes across multiple cell and tissue types by a new affinity capture method

Centrosomes are the major microtubule-organising centers in animals and play fundamental roles in many cellular processes. Understanding how their composition varies across diverse cell types and how it is altered in disease aremajor unresolved questions, yet currently available centrosome isolation protocols are cumbersome, time-consuming and lack scalability. Here, we report the development of centrosome affinity capture (CAPture)-mass spectrometry (MS), a powerful one-step method to obtain high-resolution centrosome proteomes from untransformed and primary cell lines. Utilising a synthetic peptide derived from CCDC61 protein, CAPture specifically isolates intact centrosomes. Importantly, as a bead-based affinity method, it enables rapid sample processing and multiplexing unlike conventional approaches. Our study demonstrates the power of CAPture-MS to elucidate cell typedependent heterogeneity in centrosome composition, dissect hierarchical interactions and identify novel components. Overall, CAPture-MS represents a transformative tool to unveil temporal, regulatory, cell type- and tissue-specific changes in centrosome proteomes in health and disease.