title: Monomeric fluorescent timers that change color from blue to red report on cellular trafficking.
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 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.
authors.affiliation: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
authors.forename: Fedor V
authors.initials: FV
authors.lastname: Subach
authors.forename: Oksana M
authors.initials: OM
authors.lastname: Subach
authors.forename: Illia S
authors.initials: IS
authors.lastname: Gundorov
authors.forename: Kateryna S
authors.initials: KS
authors.lastname: Morozova
authors.forename: Kiryl D
authors.initials: KD
authors.lastname: Piatkevich
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Cuervo
authors.forename: Vladislav V
authors.initials: VV
authors.lastname: Verkhusha
mesh_terms: D001692:Biological Transport; D003116:Color; D005453:Fluorescence; D006358:Hot Temperature