Tracking protein aggregation and mislocalization in cells with flow cytometry

YM Ramdzan; S Polling; CPZ Chia; IHW Ng; AR Ormsby; NP Croft; AW Purcell; MA Bogoyevitch; DCH Ng; PA Gleeson; DM Hatters

Journal article

Tracking protein aggregation and mislocalization in cells with flow cytometry

YM Ramdzan, S Polling, CPZ Chia, IHW Ng, AR Ormsby, NP Croft, AW Purcell, MA Bogoyevitch, DCH Ng, PA Gleeson, DM Hatters

Nature Methods | Published : 2012

DOI: 10.1038/nmeth.1930

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Abstract

We applied pulse-shape analysis (PulSA) to monitor protein localization changes in mammalian cells by flow cytometry. PulSA enabled high-throughput tracking of protein aggregation, translocation from the cytoplasm to the nucleus and trafficking from the plasma membrane to the Golgi as well as stress-granule formation. Combining PulSA with tetracysteine-based oligomer sensors in a cell model of Huntington's disease enabled further separation of cells enriched with monomers, oligomers and inclusion bodies. © 2012 Nature America, Inc. All rights reserved.

University of Melbourne Researchers

Paul Gleeson Author

Danny M Hatters Author

Related Projects (1)

Understanding how misshapen proteins accumulate and kill cells during Huntington's Disease

Mutations in the huntingtin gene cause Huntington's disease by making the gene product aggregate together into non-normal and different size..

Grants

Awarded by National Health and Medical Research Council

Funding Acknowledgements

This work was funded by grants to D. M. H. (Australian National Health and Medical Research Council project grant 566640). D. M. H. is funded as a Grimwade fellow, supported by the Miegunyah Trust. D.C.H.N. is funded as a CR Roper fellow. The SOD1-GFP fusion constructs were provided by B.J. Turner (Florey Neurosciences Institute, Australia). HSPA1A (Hsp70) and DNAJB1 (Hsp40) in the pCMV vector was provided by P. Muchowski (Gladstone Institute of Neurological Disease and University of California, San Francisco).