@article {47685,
	title = {Bright monomeric photoactivatable red fluorescent protein for two-color super-resolution sptPALM of live cells.},
	journal = {Journal of the American Chemical Society},
	volume = {132},
	year = {2010},
	month = {2010 May 12},
	pages = {6481-91},
	abstract = {<p>Rapidly emerging techniques of super-resolution single-molecule microscopy of living cells rely on the continued development of genetically encoded photoactivatable fluorescent proteins. On the basis of monomeric TagRFP, we have developed a photoactivatable TagRFP protein that is initially dark but becomes red fluorescent after violet light irradiation. Compared to other monomeric dark-to-red photoactivatable proteins including PAmCherry, PATagRFP has substantially higher molecular brightness, better pH stability, substantially less sensitivity to blue light, and better photostability in both ensemble and single-molecule modes. Spectroscopic analysis suggests that PATagRFP photoactivation is a two-step photochemical process involving sequential one-photon absorbance by two distinct chromophore forms. True monomeric behavior, absence of green fluorescence, and single-molecule performance in live cells make PATagRFP an excellent protein tag for two-color imaging techniques, including conventional diffraction-limited photoactivation microscopy, super-resolution photoactivated localization microscopy (PALM), and single particle tracking PALM (sptPALM) of living cells. Two-color sptPALM imaging was demonstrated using several PATagRFP tagged transmembrane proteins together with PAGFP-tagged clathrin light chain. Analysis of the resulting sptPALM images revealed that single-molecule transmembrane proteins, which are internalized into a cell via endocytosis, colocalize in space and time with plasma membrane domains enriched in clathrin light-chain molecules.</p>
},
	keywords = {Amino Acid Sequence, Animals, Cell Survival, Cercopithecus aethiops, Color, COS Cells, Hela Cells, Humans, Light, Luminescent Proteins, Membrane Proteins, Microscopy, Molecular Imaging, Molecular Sequence Data, Mutagenesis, Photochemical Processes, Temperature},
	issn = {1520-5126},
	doi = {10.1021/ja100906g},
	author = {Subach, Fedor V and Patterson, George H and Renz, Malte and Lippincott-Schwartz, Jennifer and Verkhusha, Vladislav V}
}
