Expression and optical properties of green fluorescent protein expressed in different cellular environments
Expression and optical properties of green fluorescent protein expressed in different cellular environments
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DOI：
10.1016/j.jbiotec.2005.04.001
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发表时间：
2005-10-10
影响因子：
4.1
通讯作者：
Yang, JJ
中科院分区：
文献类型：
--
作者：
Zou, J;Ye, YM;Yang, JJ
本研究调查了绿色荧光蛋白（（;FP））变体在HeLa细胞的胞质溶胶和内质网（ER）中的表达，并评估了不同细胞环境对这些GFP变体的荧光特性的影响。已经通过添加不同的N-或C-末端信号序列构建了几种GFP变体。这些蛋白质在HeLa细胞中的不同细胞区室中表达和折叠。通过共聚焦显微镜评估DsRed 2-ER的共定位来确认这些GFP变体靶向内质网的定位。将靶向GFP变体的信号肽添加到ER或胞质溶胶中似乎不会改变光谱或这些GFP变体。然而，这些GFP变体在ER中的荧光强度显著低于在胞质溶胶中的荧光强度。因此，结果清楚地表明，细胞环境影响体内绿色荧光蛋白的形成和/或成熟。这些发现将有助于在未来的发展和应用的GFP技术，旨在调查在ER和胞质溶胶中执行的细胞功能。(C)2005 Elsevier B. V.保留所有权利。
This study has investigated the expression of green fluorescent protein ((;FP) variants in the cytosol and the endoplasmic reticulum (ER) of HeLa cells and evaluated the effects of the different cellular environments on the fluorescence properties of these GFP variants. Several GFP variants have been constructed by adding, different N- or C-terminal signal sequences, These proteins were expressed and folded in distinct cellular compartments in HeLa cells. The localization of these GFP variants targeted to the endoplasmic recticulum was confirmed by the co-localization of DsRed2-ER as assessed by confocal microscopy. The addition of signal peptides targeting GFP variants to the ER or cytosol did not appear to alter the optical spectra or these GFP variants. However, the fluorescence intensity of these GFP variants in the ER was significantly less than that in (tic cytosol. Thus, the results clearly suggest that the cellular environment affects the formation and/or maturation of green fluorescence protein in vivo. These findings will be helpful in the future development and application of GFP technology aimed at investigating cellular functions performed in the ER and the cytosol. (C) 2005 Elsevier B.V. All rights reserved.