Quantification of photosensitized singlet oxygen production by a fluorescent protein

Xavier Ragàs, Laurie P. Cooper, John H. White, Santi Nonell, Cristina Flors

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Resum

Fluorescent proteins are increasingly becoming actuators in a range of cell biology techniques. One of those techniques is chromophore-assisted laser inactivation (CALI), which is employed to specifically inactivate the function of target proteins or organelles by producing photochemical damage. CALI is achieved by the irradiation of dyes that are able to produce reactive oxygen species (ROS). The combination of CALI and the labelling specificity that fluorescent proteins provide is useful to avoid uncontrolled photodamage, although the inactivation mechanisms by ROS are dependent on the fluorescent protein and are not fully understood. Herein, we present a quantitative study of the ability of the red fluorescent protein TagRFP to produce ROS, in particular singlet oxygen (1O2). TagRFP is able to photosensitize 1O2 with an estimated quantum yield of 0.004. This is the first estimation of a quantum yield of 1O2 production value for a GFP-like protein. We also find that TagRFP has a short triplet lifetime compared to EGFP, which reflects relatively high oxygen accessibility to the chromophore. The insight into the structural and photophysical properties of TagRFP has implications in improving fluorescent proteins for fluorescence microscopy and CALI.

Idioma originalAnglès
Pàgines (de-a)161-165
Nombre de pàgines5
RevistaChemPhysChem
Volum12
Número1
DOIs
Estat de la publicacióPublicada - 17 de gen. 2011

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