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Why Green Fluorescent Fusion Proteins Have Not Been Observed in the Vacuoles of Higher Plants by Kentaro Tamura; Tomoo Shimada; Eiichiro Ono; Yoshikazu Tanaka; Akira Nagatani; Sho-ich Higashi; Masakatsu Watanabe; Mikio Nishimura; Ikuko Hara-Nishimura is a Agricultural and Biological Sciences article available to read on EtoBox.
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## Summary Green fluorescent protein (GFP) makes it possible for organelles and protein transport pathways to be visualized in living cells. However, GFP fluorescence has not yet been observed in the vacuoles of any organs of higher plants. We found that the fluorescence of a vacuole‐targeted GFP was stably observed in the vacuoles of transgenic __Arabidopsis__ plants under dark conditions, and that the fluorescence rapidly disappeared under light conditions. The vacuolar GFP was rapidly degraded within 1 h in the light, especially blue light. An inhibitor of vacuolar type H^+^‐ATPase, concanamycin A, and an inhibitor of papain‐type cysteine proteinase, E‐64d, abolished both the light‐dependent disappearance of GFP fluorescence and GFP degradation in the vacuoles. An __in vitro__ assay showed that bacterially expressed GFP was degraded by extracts of __Arabidopsis__ cultured‐cell protoplasts at an acidic pH in the light. These results suggest that blue light induced a conformational change in GFP, and the resulting GFP in the vacuole was easily degraded by vacuolar papain‐type cysteine proteinase(s) under the acidic pH. The light‐dependent degradation accounts for the failure to ob
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- Author
- Kentaro Tamura; Tomoo Shimada; Eiichiro Ono; Yoshikazu Tanaka; Akira Nagatani; Sho-ich Higashi; Masakatsu Watanabe; Mikio Nishimura; Ikuko Hara-Nishimura
- Publisher
- John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0960-7412)
- Published
- 2003
- Language
- EN
- Field
- Agricultural and Biological Sciences (Life Sciences)