A general and efficient method for the site-specific dual-labeling of proteins for single molecule fluorescence resonance energy transfer.
A general and efficient method for the site-specific dual-labeling of proteins for single molecule fluorescence resonance energy transfer.
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DOI:
10.1021/ja807430h
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发表时间:
2008-12-31
影响因子:
15
通讯作者:
Deniz AA
中科院分区:
文献类型:
--
作者:
Brustad EM;Lemke EA;Schultz PG;Deniz AA
A general strategy for the site-specific dual-labeling of proteins for single-molecule fluorescence resonance energy transfer (smFRET) is presented. A genetically encoded unnatural ketone amino acid was labeled with a hydroxylamine-containing fluorophore with high yield (>95%) and specificity. This methodology was used to construct dual-labeled T4 lysozyme variants, allowing the study of T4 lysozyme folding at single-molecule resolution. The presented strategy is anticipated to expand the scope of single-molecule protein structure and function studies.
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