pH敏感型荧光蛋白及其在植物细胞生物学中的应用

doi:10.3724/SP.J.1259.2015.00394

植物学报 ›› 2015, Vol. 50 ›› Issue (3): 394-404.DOI: 10.3724/SP.J.1259.2015.00394

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pH敏感型荧光蛋白及其在植物细胞生物学中的应用

薛轶群¹, 宋凯¹, 范路生¹, 万迎朗², 林金星^1,^2,^*()

¹中国科学院植物研究所, 北京 100093
²北京林业大学生物科学与技术学院, 北京 100083

收稿日期:2014-06-04 接受日期:2014-09-09 出版日期:2015-05-01 发布日期:2015-04-08
通讯作者: 林金星
作者简介:
? 共同第一作者
基金资助:
国家重点基础研究发展规划(No.2011CB809103)、国家自然科学基金创新研究群体(No.31121065)、北京林业大学杰出青年人才培育计划(No.JC2013-2)、新世纪优秀人才计划(No.NECT-12-0785)和国家自然科学基金(No.31271433)

pH-sensitive Fluorescent Proteins and Their Applications in Plant Cell Biology

Yiqun Xue¹, Kai Song¹, Lusheng Fan¹, Yinglang Wan², Jinxing Lin^{1, 2, *}

¹Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
²College of Biological Sciences and Technology, Beijing Forestry University, Beijing 100083, China

Received:2014-06-04 Accepted:2014-09-09 Online:2015-05-01 Published:2015-04-08
Contact: Lin Jinxing
About author:
? These authors contributed equally to this paper

摘要/Abstract

摘要： pH敏感型荧光蛋白, 即pHluorin, 是荧光强度及光谱特征随环境pH值的变化而改变的一类荧光蛋白。人们通过对密码子使用偏好和特定剪切位点的修饰, 已使pHluorin及其衍生物成功地在动物、植物和真菌细胞中正常表达, 为测量细胞内微环境pH值的变化, 并研究活细胞内依赖或导致pH变化的生理过程提供了有力工具。该文总结了目前已报道的pH敏感型荧光蛋白的种类及特性, 并对其在细胞生物学, 特别是植物细胞生物学中的应用进行了详细介绍。随着报告基因技术及检测方法的不断改进, pHluorin将在植物科学领域发挥更大的作用。

Abstract: pHluorin is a pH-sensitive variant of green fluorescent protein; its fluorescence signals display a strong pH dependence. Since the codon usage was optimized and the site of aberrant splicing was removed, pHluorin and its derivatives have been successfully expressed and used in animal, plant and fungi cells. These pH sensors have helped shed light on measure of the difference in pH of cellular microenvironment and many cell functions for which intracellular pH is an important modulator. Here, we review the characteristics of known pHluorins and their applications in cell biology, particularly plant cell biology. With the improvements in pHluorins by genetic engineering and the advancing in detection technology, the future for pHluorins in plant science is exciting.

薛轶群, 宋凯, 范路生, 万迎朗, 林金星. pH敏感型荧光蛋白及其在植物细胞生物学中的应用. 植物学报, 2015, 50(3): 394-404.

Yiqun Xue, Kai Song, Lusheng Fan, Yinglang Wan, Jinxing Lin. pH-sensitive Fluorescent Proteins and Their Applications in Plant Cell Biology. Chinese Bulletin of Botany, 2015, 50(3): 394-404.

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荧光蛋白名称	类型	激发光(nm)	发射光(nm)	pH范围	pKa	目前应用范畴	参考文献
pHluorin	比率	395/475	510	5.4-8.4	6.9	测量动物细胞内涵体和TGN中pH值	Miesenböck et al., 1998
pHluorin2	比率	395/475	510	5.4-8.4	6.9	检测动物细胞中激素受体内吞	Mahon, 2011
RaVC	比率	395/475	510	5.4-8.4	6.9	测量曲霉菌中胞质pH值	Bagar et al., 2009
pHGFP	比率	395/475	510	5.4-8.4	6.9	测量拟南芥根中胞质pH值	Moseyko and Feldman, 2001
Ratiometric AtpHluorin	比率	395/475	510	5.4-8.4	6.9	测量拟南芥根质外体和细胞质中pH值	Gao et al., 2004
PRpHluorin	比率	395/475	510	5.4-8.4	6.9	测量拟南芥原生质体中细胞器pH值	Shen et al., 2013
Ecliptic pHluorin	盈缺	395/477	510	6.5-8.0	7.2	检测动物细胞突触囊泡胞吐	Miesenböck et al., 1998
Ecliptic AtpHluorin	盈缺	395/477	510	6.5-8.0	7.2	测量拟南芥根质外体和细胞质中pH值	Gao et al., 2004
PEpHluorin	盈缺	395/477	510	6.5-8.0	7.2	测量拟南芥原生质体中细胞器pH值	Shen et al., 2013
Superecliptic pHluorin	盈缺	477	510	5.5-8.5	7.2	检测动物细胞突触末端胞吞胞吐	Sankaranarayanan and Ryan, 2001
deGFP	比率	396	460/515	6-9.0	7.3	测量动物细胞中pH值	Hanson et al., 2002
E²GFP	比率	458/488	500/560	5-8.5	6.9/7.5	测量动物细胞中pH值	Bizzarri et al., 2006; Arosio et al., 2007
ptGFP	比率	390/475	540	3.8-8.2	7.3	测量拟南芥根质外体和细胞质中pH值	Schulte et al., 2006
pHlameleon 6	比率	458	481/533	5-7.5	5.6	基于FRET原理测量动物细胞中pH值	Esposito et al., 2008
pHusion	比率	488/458	510/600	4.5-8	5.8	基于FRET原理测量拟南芥叶肉和根的固定细胞中质外体和胞质pH值	Gjetting et al., 2012
pHlash	比率	不适用	475/525	5.4-9	不适用	基于BRET原理测量动物细胞中胞质 pH值	Zhang et al., 2012

荧光蛋白名称	类型	激发光(nm)	发射光(nm)	pH范围	pKa	目前应用范畴	参考文献
pHluorin	比率	395/475	510	5.4-8.4	6.9	测量动物细胞内涵体和TGN中pH值	Miesenböck et al., 1998
pHluorin2	比率	395/475	510	5.4-8.4	6.9	检测动物细胞中激素受体内吞	Mahon, 2011
RaVC	比率	395/475	510	5.4-8.4	6.9	测量曲霉菌中胞质pH值	Bagar et al., 2009
pHGFP	比率	395/475	510	5.4-8.4	6.9	测量拟南芥根中胞质pH值	Moseyko and Feldman, 2001
Ratiometric AtpHluorin	比率	395/475	510	5.4-8.4	6.9	测量拟南芥根质外体和细胞质中pH值	Gao et al., 2004
PRpHluorin	比率	395/475	510	5.4-8.4	6.9	测量拟南芥原生质体中细胞器pH值	Shen et al., 2013
Ecliptic pHluorin	盈缺	395/477	510	6.5-8.0	7.2	检测动物细胞突触囊泡胞吐	Miesenböck et al., 1998
Ecliptic AtpHluorin	盈缺	395/477	510	6.5-8.0	7.2	测量拟南芥根质外体和细胞质中pH值	Gao et al., 2004
PEpHluorin	盈缺	395/477	510	6.5-8.0	7.2	测量拟南芥原生质体中细胞器pH值	Shen et al., 2013
Superecliptic pHluorin	盈缺	477	510	5.5-8.5	7.2	检测动物细胞突触末端胞吞胞吐	Sankaranarayanan and Ryan, 2001
deGFP	比率	396	460/515	6-9.0	7.3	测量动物细胞中pH值	Hanson et al., 2002
E²GFP	比率	458/488	500/560	5-8.5	6.9/7.5	测量动物细胞中pH值	Bizzarri et al., 2006; Arosio et al., 2007
ptGFP	比率	390/475	540	3.8-8.2	7.3	测量拟南芥根质外体和细胞质中pH值	Schulte et al., 2006
pHlameleon 6	比率	458	481/533	5-7.5	5.6	基于FRET原理测量动物细胞中pH值	Esposito et al., 2008
pHusion	比率	488/458	510/600	4.5-8	5.8	基于FRET原理测量拟南芥叶肉和根的固定细胞中质外体和胞质pH值	Gjetting et al., 2012
pHlash	比率	不适用	475/525	5.4-9	不适用	基于BRET原理测量动物细胞中胞质 pH值	Zhang et al., 2012

pH敏感型荧光蛋白及其在植物细胞生物学中的应用

pH-sensitive Fluorescent Proteins and Their Applications in Plant Cell Biology

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