RFP tags for labeling secretory pathway proteins

被引：9

作者：

Han, Liyang ^{[1
]}

Zhao, Yanhua ^{[1
]}

Zhang, Xi ^{[3
]}

Peng, Jianxin ^{[3
]}

Xu, Pingyong ^{[2
]}

Huan, Shuangyan ^{[1
]}

Zhang, Mingshu ^{[2
]}

机构：

[1] Hunan Univ, Coll Chem & Chem Engn, State Key Lab Chemo Biosensing & Chemometr, Changsha 410082, Hunan, Peoples R China

[2] Chinese Acad Sci, Inst Biophys, Key Lab Interdisciplinary Res, Beijing 100101, Peoples R China

[3] Cent China Normal Univ, Coll Life Sci, Wuhan 430079, Hubei, Peoples R China

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2014年 / 447卷 / 03期

基金：

中国国家自然科学基金; 北京市自然科学基金;

关键词：

Red fluorescent proteins; Artificial puncta; K-d; Evaluation method; pKa; GREEN FLUORESCENT PROTEIN; BRIGHT; MEMBRANE;

D O I：

10.1016/j.bbrc.2014.04.013

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Red fluorescent proteins (RFPs) are useful tools for live cell and multi-color imaging in biological studies. However, when labeling proteins in secretory pathway, many RFPs are prone to form artificial puncta, which may severely impede their further uses. Here we report a fast and easy method to evaluate RFPs fusion properties by attaching RFPs to an environment sensitive membrane protein Orai1. In addition, we revealed that intracellular artificial puncta are actually colocalized with lysosome, thus besides monomeric properties, pica value of RFPs is also a key factor for forming intracellular artificial puncta. In summary, our current study provides a useful guide for choosing appropriate RFP for labeling secretory membrane proteins. Among RFPs tested, mOrange2 is highly recommended based on excellent monomeric property, appropriate pKa and high brightness. (C) 2014 Elsevier Inc. All rights reserved.

引用

页码：508 / 512

页数：5

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