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

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.

Pubmed ID: 19136976

Authors

  • Subach FV
  • Subach OM
  • Gundorov IS
  • Morozova KS
  • Piatkevich KD
  • Cuervo AM
  • Verkhusha VV

Journal

Nature chemical biology

Publication Data

February 16, 2009

Associated Grants

  • Agency: NIGMS NIH HHS, Id: GM070358
  • Agency: NIGMS NIH HHS, Id: GM073913
  • Agency: NIA NIH HHS, Id: R01 AG021904
  • Agency: NIA NIH HHS, Id: R01 AG021904-06

Mesh Terms

  • Biological Transport
  • Color
  • Fluorescence
  • Hot Temperature