Journal article
Conformation Sensors that Distinguish Monomeric Proteins from Oligomers in Live Cells
YM Ramdzan, RM Nisbet, J Miller, S Finkbeiner, AF Hill, DM Hatters
Chemistry and Biology | CELL PRESS | Published : 2010
Abstract
Proteins prone to misfolding form large macroscopic deposits in many neurodegenerative diseases. Yet the in situ aggregation kinetics remains poorly understood because of an inability to demarcate precursor oligomers from monomers. We developed a strategy for mapping the localization of soluble oligomers and monomers directly in live cells. Sensors for mutant huntingtin, which forms aggregates in Huntington's disease, were made by introducing a tetracysteine motif into huntingtin that becomes occluded from binding biarsenical fluorophores in oligomers, but not monomers. Up to 70% of the diffusely distributed huntingtin molecules appeared as submicroscopic oligomers in individual neuroblastom..
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Awarded by National Institute of Neurological Disorders and Stroke
Funding Acknowledgements
We thank K. Goldie for performing the electron microscopy experiments. This work was funded by grants to A.F.H. (National Health and Medical Research Council program grant 400202 and Career Development Award [Level 2] 509400) and D.M.H. (National Health and Medical Research Council project grant 566640). D.M.H. is a CR Roper Fellow. S.F. was supported by the National Institutes of Neurological Disease and Stroke (R01 2NS039074 and R01 2NS045191), the CHDI Foundation, and the Taube-Koret Center for Huntington's Disease Research, J.M. is supported by a predoctoral fellowship from the Hillbloom Foundation as well as the NIH-NIGMS UCSF Medical Scientist Training Program.