期刊
BIOPHYSICAL JOURNAL
卷 98, 期 12, 页码 3078-3085出版社
CELL PRESS
DOI: 10.1016/j.bpj.2010.02.058
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类别
资金
- Cell Migration Consortium [U54 GM064346]
- National Institutes of Health [P41-RRO3155, P50-GM076516, NS045283]
- Cancer Center [CA-62203]
Protein aggregation is a hallmark of several neurodegenerative diseases including Huntington's disease. We describe the use of the recently developed number and brightness method (N&B) that uses confocal images to monitor aggregation of Huntingtin exon 1 protein (Httex1p) directly in living cells. N&B measures the molecular brightness of protein aggregates in the entire cell noninvasively based on intensity fluctuations at each pixel in an image. N&B applied to mutant Httex1 p in living cells showed a two-step pathway leading to inclusion formation that is polyQ length dependent and involves four phases. An initial phase of monomer accumulation is followed by formation of small oligomers (5-15 proteins); as protein concentration increases, an inclusion is seeded and forms in the cytoplasm; the growing inclusion recruits most of the Httex1 p and depletes the cell leaving only a low concentration of monomers. The behavior of Httex1 p in COS-7 and ST14A cells is compared.
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