网站地图 | 联系我们 | 管理系统 | 登录邮箱 | 内网办公 | English | 中国科学院    
首页 所况介绍 机构设置 科研成果 研究队伍 合作交流 研究生教育 党群园地 科学传播
 
 
 当前位置:首页 >>> 论文库

Single-Molecule Imaging and Computational Microscopy Approaches Clarify the Mechanism of the Dimerization and Membrane Interactions of Green Fluorescent Protein

作 者:Wang XH, Song K, Li Y, Tang L, Deng X*
影响因子: 3.687
刊物名称: International Journal of Molecular Sciences
出版年份: 2019
卷: 20  期: 6   页码: 1410

文章摘要 : 

Green fluorescent protein (GFP) is widely used as a biomarker in living systems, however GFP and its variants are prone to forming low-affinity dimers under physiological conditions. This undesirable tendency is exacerbated when fluorescent proteins (FP) are confined to membranes fused to naturally-oligomeric proteins or expressed at high levels in cells. Oligomerization of FPs introduces artifacts into the measurement of subunit stoichiometry as well as interactions between proteins fused to FPs. Introduction of a single mutation A206K has been shown to disrupt hydrophobic interactions in the region responsible for GFP dimerization thereby contributing to its monomerization. Nevertheless a detailed understanding of how this single amino acid-dependent inhibition of dimerization in GFP occurs at the atomic level is still lacking. Single-molecule experiments combined with computational microscopy (atomistic molecular dynamics) revealed that the amino group of A206 contributes to GFP dimer formation via a multivalent electrostatic interaction. We further showed that myristoyl modification is an efficient mechanism to promote membrane attachment of GFP. Molecular dynamics-based site-directed mutagenesis has been used to identify the key functional residues in FPs. The data presented here have been utilized as a monomeric control in downstream single-molecule studies facilitating more accurate stoichiometry quantification of functional protein complexes in living cells.


| 本站导航 | 园区风光| 联系我们 |
版权所有:中国科学院植物研究所 地址:北京市海淀区香山南辛村20号 邮编:100093 电话:010-62590835
网站备案号:京ICP备05002828号       文保网安备案号:1101080078