Abstract
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°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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 118-126 |
| Number of pages | 9 |
| Journal | Nature Chemical Biology |
| Volume | 5 |
| Issue number | 2 |
| DOIs | |
| State | Published - Feb 6 2009 |
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ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
Cite this
Monomeric fluorescent timers that change color from blue to red report on cellular trafficking. / Subach, Fedor V.; Subach, Oksana M.; Gundorov, Illia S.; Morozova, Kateryna S.; Piatkevich, Kiryl D.; Cuervo, Ana Maria; Verkhusha, Vladislav V.
In: Nature Chemical Biology, Vol. 5, No. 2, 06.02.2009, p. 118-126.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Monomeric fluorescent timers that change color from blue to red report on cellular trafficking
AU - Subach, Fedor V.
AU - Subach, Oksana M.
AU - Gundorov, Illia S.
AU - Morozova, Kateryna S.
AU - Piatkevich, Kiryl D.
AU - Cuervo, Ana Maria
AU - Verkhusha, Vladislav V.
PY - 2009/2/6
Y1 - 2009/2/6
N2 - 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°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.
AB - 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°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.
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U2 - 10.1038/nchembio.138
DO - 10.1038/nchembio.138
M3 - Article
C2 - 19136976
AN - SCOPUS:58249123661
VL - 5
SP - 118
EP - 126
JO - Nature Chemical Biology
JF - Nature Chemical Biology
SN - 1552-4450
IS - 2
ER -