Photoactivation turns green fluorescent protein red.
In the few years since its gene was first cloned, the Aequorea victoria green fluorescent protein (GFP) has become a powerful tool in cell biology, functioning as a marker for gene expression, protein localization and protein dynamics in living cells. GFP variants with improved fluorescence intensity and altered spectral characteristics have been identified, but additional GFP variants are still desirable for multiple labeling experiments, protein interaction studies and improved visibility in some organisms. In particular, long-wavelength (red) fluorescence has remained elusive. Here we describe a red-emitting, green-absorbing fluorescent state of GFP that is generated by photoactivation with blue light. GFP can be switched to its red-emitting state easily with a laser or fluorescence microscope lamp under conditions of low oxygen concentration. This previously unnoticed ability enables regional, non-invasive marking of proteins in vivo. In particular, we report here the use of GFP photoactivation to make the first direct measurements of protein diffusion in the cytoplasm of living bacteria.
Elowitz
M B
MB
Department of Physics, Princeton University, New Jersey 08544, USA.
Surette
M G
MG
Wolf
P E
PE
Stock
J
J
Leibler
S
S
D004926:Escherichia coli / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008027:Light; D008164:Luminescent Proteins / Q000528:radiation effects*; D010100:Oxygen / Q000378:metabolism; D010777:Photochemistry
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D010100:Oxygen