Monomeric fluorescent timers that change color from blue to red report on cellular trafficking

Fedor V. Subach, Oksana M. Subach, Illia S. Gundorov, Kateryna S. Morozova, Kiryl D. Piatkevich, Ana Maria Cuervo, Vladislav V. Verkhusha

Research output: Contribution to journalArticle

86 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)118-126
Number of pages9
JournalNature Chemical Biology
Volume5
Issue number2
DOIs
StatePublished - Feb 6 2009

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Lysosome-Associated Membrane Glycoproteins
Color
Fluorescence
Protein Transport
Endosomes
Lysosomes
Insects
Cell Membrane
Bacteria
Temperature

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 journalArticle

Subach, Fedor V. ; Subach, Oksana M. ; Gundorov, Illia S. ; Morozova, Kateryna S. ; Piatkevich, Kiryl D. ; Cuervo, Ana Maria ; Verkhusha, Vladislav V. / Monomeric fluorescent timers that change color from blue to red report on cellular trafficking. In: Nature Chemical Biology. 2009 ; Vol. 5, No. 2. pp. 118-126.
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