Folding latency of fluorescent proteins affects the mitochondrial localization of fusion protein
The discovery of fluorescent proteins (FPs) has revolutionized cell biology. The fusion of targeting sequences to FPs enables the investigation of cellular organelles and their dynamics; however, occasionally, such fluorescent fusion proteins (FFPs) exhibit behavior different from that of the native proteins. Here, we constructed a color pallet comprising different organelle markers and found that FFPs targeted to the mitochondria were mislocalized when fused to certain types of FPs. Such FPs included several variants of Aequorea victoria green FP (aqGFP) and a monomeric variant of the red FP. Because the FFPs that are mislocalized include FPs with faster maturing or folding mutations, the increase in the maturation rate is likely to prevent their expected localization. Indeed, when we reintroduced amino acid substitutions so that the FP sequences were equivalent to that of wild-type aqGFP, FFP localization to the mitochondria was significantly enhanced. Moreover, similar amino acid substitutions improved the localization of mitochondria-targeted pHluorin, which is a pH-sensitive variant of GFP, and its capability to monitor pH changes in the mitochondrial matrix. Our findings demonstrate the importance of selecting FPs that maximize FFP function.
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Sayaka Kashiwagi (add twitter)
Yoichiro Fujioka (add twitter)
Aya O Satoh (edit)
Aiko Yoshida (edit)
Mari Fujioka (add twitter)
Prabha Nepal (edit)
Atsuhi Tsuzuki (add twitter)
Ozorea Aoki (add twitter)
Sarad Paudel (add twitter)
Hitoshi Sasajima (add twitter)
Yusuke Ohba (add twitter)
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Cell Biology

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09/13/19 02:02PM
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biorxiv_cellbio: Folding latency of fluorescent proteins affects the mitochondrial localization of fusion protein https://t.co/qYPTDj0YJI #biorxiv_cellbio
biorxivpreprint: Folding latency of fluorescent proteins affects the mitochondrial localization of fusion protein https://t.co/4ebyUcQ2Li #bioRxiv
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