期刊
NATURE NEUROSCIENCE
卷 13, 期 7, 页码 897-U149出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nn.2580
关键词
-
资金
- German Academy for Natural Scientists Leopoldina
- US National Institutes of Health
- Howard Hughes Medical Institute
- Ministere de l'Enseignement Superieur et de la Recherche
- Nationale de la Recherche MorphoSynDiff-INSERM
Protein translation has been implicated in different forms of synaptic plasticity, but direct in situ visualization of new proteins is limited to one or two proteins at a time. Here we describe a metabolic labeling approach based on incorporation of noncanonical amino acids into proteins followed by chemoselective fluorescence tagging by means of 'click chemistry'. After a brief incubation with azidohomoalanine or homopropargylglycine, a robust fluorescent signal was detected in somata and dendrites. Pulse-chase application of azidohomoalanine and homopropargylglycine allowed visualization of proteins synthesized in two sequential time periods. This technique can be used to detect changes in protein synthesis and to evaluate the fate of proteins synthesized in different cellular compartments. Moreover, using strain-promoted cycloaddition, we explored the dynamics of newly synthesized membrane proteins using single-particle tracking and quantum dots. The newly synthesized proteins showed a broad range of diffusive behaviors, as would be expected for a pool of labeled proteins that is heterogeneous.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据