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【bio-news】化学生物学：荧光计时器
New fluorescent 'timers' that gradually change colour from blue to red could allow researchers to track the age and dynamic behaviour of proteins in living cells.
Previous work suggested that some red fluorescent proteins start out fluorescing blue, but then change to red as the protein is chemically modified over time. Vladislav Verkhusha and his colleagues at the Albert Einstein College of Medicine in New York mutated a red fluorescent protein called mCherry, then screened for mutants that had altered maturation rates from blue to red.
The researchers developed three fluorescent proteins, each with a specific maturation rate. The proteins were used to track newly synthesized proteins in mammalian cells grown in culture.
附录一 原文
Subach FV, Subach OM, Gundorov IS, Morozova KS, Piatkevich KD, Cuervo AM, Verkhusha VV. Monomeric fluorescent timers that change color from blue to red report on cellular trafficking.Nat Chem Biol. 2009 Jan 11. [Epub ahead of print]
Based on the mechanism for chromophore formation in red fluorescent proteins, we developed three mCherry-derived monomeric variants, called fluorescent timers (FTs), that change their fluorescence from the blue to red over time. These variants exhibit distinctive fast, medium and slow blue-to-red chromophore maturation rates that depend on the temperature. At 37 degrees C, the maxima of the blue fluorescence are observed at 0.25, 1.2 and 9.8 h for the purified fast-FT, medium-FT and slow-FT, respectively. The half-maxima of the red fluorescence are reached at 7.1, 3.9 and 28 h, respectively. The FTs show similar timing behavior in bacteria, insect and mammalian cells. Medium-FT allowed for tracking of the intracellular dynamics of the lysosome-associated membrane protein type 2A (LAMP-2A) and determination of its age in the targeted compartments. The results indicate that LAMP-2A transport through the plasma membrane and early or recycling endosomes to lysosomes is a major pathway for LAMP-2A trafficking. [标签:content1][标签:content2]
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作者:admin@医学,生命科学 2011-08-23 17:33
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