Direct Proximity Tagging of Small Molecule Protein Targets Using an Engineered NEDD8 Ligase
Hill, Zachary B.1; Pollock, Samuel B.1; Zhuang, Min1,3; Wells, James A.1,2
2016-10-12
发表期刊JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
ISSN0002-7863
卷号138期号:40页码:13123-13126
发表状态已发表
DOI10.1021/jacs.6b06828
摘要Identifying the protein targets of bioactive small molecules remains a major problem in the discovery of new chemical probes and therapeutics. While activity based probes and photo-cross-linkers have had success in identifying protein targets of small molecules, each technique has limitations. Here we describe a method for direct proximity tagging of proteins that bind small molecules. We engineered a promiscuous ligase based on the NEDD8 conjugating enzyme, Ubc12, which can be covalently linked to a small molecule of interest. When target proteins bind the small molecule, they are directly labeled on surface lysines with a biotinylated derivative of the small ubiquitin homologue, NEDD8. This unique covalent tag can then be used to identify the small molecule binding proteins. Utilizing the drug dasatinib, we have shown that dasatinib-directed NEDDylation occurs for known endogenous protein binders in complex cell lysates. In addition, we have been able to improve NEDDylation efficiency through rational mutagenesis. Finally, we have shown that affinity-directed NEDDylation can be applied to two other protein ligand interactions beyond kinases. Proximity tagging using this engineered ligase requires direct binding of the target and, thus, provides a useful and orthogonal approach to facilitate small molecule target identification.
收录类别SCI ; EI
语种英语
资助项目NIH[R01 CA191018]
WOS研究方向Chemistry
WOS类目Chemistry, Multidisciplinary
WOS记录号WOS:000385469600007
出版者AMER CHEMICAL SOC
EI入藏号20164302935931
EI主题词Amino acids ; Bins ; Enzymes ; Molecules ; Probes
EI分类号Storage:694.4 ; Organic Compounds:804.1 ; Atomic and Molecular Physics:931.3
WOS关键词KINASE ; CELLS ; INHIBITORS ; DASATINIB ; CANCER
原始文献类型Article
引用统计
文献类型期刊论文
条目标识符https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1671
专题生命科学与技术学院_PI研究组_庄敏组
通讯作者Wells, James A.
作者单位1.Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94158 USA
2.Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94158 USA
3.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China
推荐引用方式
GB/T 7714
Hill, Zachary B.,Pollock, Samuel B.,Zhuang, Min,et al. Direct Proximity Tagging of Small Molecule Protein Targets Using an Engineered NEDD8 Ligase[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY,2016,138(40):13123-13126.
APA Hill, Zachary B.,Pollock, Samuel B.,Zhuang, Min,&Wells, James A..(2016).Direct Proximity Tagging of Small Molecule Protein Targets Using an Engineered NEDD8 Ligase.JOURNAL OF THE AMERICAN CHEMICAL SOCIETY,138(40),13123-13126.
MLA Hill, Zachary B.,et al."Direct Proximity Tagging of Small Molecule Protein Targets Using an Engineered NEDD8 Ligase".JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 138.40(2016):13123-13126.
条目包含的文件 下载所有文件
文件名称/大小 文献类型 版本类型 开放类型 使用许可
个性服务
查看访问统计
谷歌学术
谷歌学术中相似的文章
[Hill, Zachary B.]的文章
[Pollock, Samuel B.]的文章
[Zhuang, Min]的文章
百度学术
百度学术中相似的文章
[Hill, Zachary B.]的文章
[Pollock, Samuel B.]的文章
[Zhuang, Min]的文章
必应学术
必应学术中相似的文章
[Hill, Zachary B.]的文章
[Pollock, Samuel B.]的文章
[Zhuang, Min]的文章
相关权益政策
暂无数据
收藏/分享
文件名: 10.1021@jacs.6b06828.pdf
格式: Adobe PDF
此文件暂不支持浏览
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。