Precision Electrophile Tagging in Caenorhabditis elegans,Biochemistry

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Precision Electrophile Tagging in Caenorhabditis elegans
Biochemistry ( IF 2.9 ) Pub Date : 2017-09-12 00:00:00 , DOI: 10.1021/acs.biochem.7b00642
Marcus J. C. Long ₁ , Daniel A. Urul ₁ , Shivansh Chawla ₁ , Hong-Yu Lin ₁ , Yi Zhao ₁ , Joseph A. Haegele ₁ , Yiran Wang ₁ , Yimon Aye _{1,

2}

Affiliation

Adduction of an electrophile to privileged sensor proteins and the resulting phenotypically dominant responses are increasingly appreciated as being essential for metazoan health. Functional similarities between the biological electrophiles and electrophilic pharmacophores commonly found in covalent drugs further fortify the translational relevance of these small-molecule signals. Genetically encodable or small-molecule-based fluorescent reporters and redox proteomics have revolutionized the observation and profiling of cellular redox states and electrophile–sensor proteins, respectively. However, precision mapping between specific redox-modified targets and specific responses has only recently begun to be addressed, and systems tractable to both genetic manipulation and on-target redox signaling in vivo remain largely limited. Here we engineer transgenic Caenorhabditis elegans expressing functional HaloTagged fusion proteins and use this system to develop a generalizable light-controlled approach to tagging a prototypical electrophile–sensor protein with native electrophiles in vivo. The method circumvents issues associated with low uptake/distribution and toxicity/promiscuity. Given the validated success of C. elegans in aging studies, this optimized platform offers a new lens with which to scrutinize how on-target electrophile signaling influences redox-dependent life span regulation.

中文翻译：

秀丽隐杆线虫的精密亲电标签

亲电子对特权传感器蛋白的加成以及由此产生的表型显性反应日益被视为对后生健康必不可少的。共价药物中常见的生物亲电试剂与亲电药效基团之间的功能相似性进一步加强了这些小分子信号的翻译相关性。遗传可编码或基于小分子的荧光报告基因和氧化还原蛋白质组学分别革新了细胞氧化还原状态和亲电传感器蛋白的观察和分析。然而，特异性氧化还原修饰的靶标和特异性应答之间的精确定位直到最近才开始被解决，并且在体内遗传控制和靶上氧化还原信号均易于处理的系统仍然受到很大限制。秀丽隐杆线虫表达功能性HaloTagged融合蛋白，并使用该系统开发出一种通用的光控方法，以在体内用天然亲电试剂标记原型亲电传感器蛋白。该方法规避了与低摄取/分布和毒性/滥交有关的问题。鉴于秀丽隐杆线虫在衰老研究中的成功验证，这个优化的平台提供了一个新的视角，可以用来研究目标亲电体信号如何影响依赖于氧化还原的寿命调节。

更新日期：2017-09-12

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