Discovery of a 29-Amino-Acid Reactive Abiotic Peptide for Selective Cysteine Arylation,ACS Chemical Biology

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Discovery of a 29-Amino-Acid Reactive Abiotic Peptide for Selective Cysteine Arylation
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2017-12-28 00:00:00 , DOI: 10.1021/acschembio.7b00520
Ethan D. Evans ₁ , Bradley L. Pentelute ₁

Affiliation

The regio- and chemoselective modification of proteins or peptides with chemical reagents is often challenging. One approach to overcome this problem involves identifying abiotic polypeptide sequences that react with specific small molecules. Toward this goal, we profiled ∼5 × 10¹³ randomized 30-mer peptides using mRNA display and high-throughput sequencing in search of polypeptides that can undergo cysteine arylation with a water-soluble perfluoroarene. Within this vast chemical space, we discovered a cysteine-containing sequence with a second-order rate constant of 0.29 M^–1 s^–1 for arylation. An N- and C-terminal truncation reduced the reaction rate, as did the addition of denaturants. When the reactive peptide was covalently fused to the enzyme Sortase A, we observed regiospecific arylation at a single cysteine site, leaving the enzyme’s active site cysteine unchanged. Taken together, these results demonstrate that long polypeptides of defined sequence, when matched with the appropriate reactive group, can be used for selective arylation of cysteine in water.

中文翻译：

发现一种用于选择性半胱氨酸酰化的29种氨基酸反应性非生物肽

用化学试剂对蛋白质或肽进行区域和化学选择性修饰通常是具有挑战性的。解决该问题的一种方法涉及鉴定与特定小分子反应的非生物多肽序列。为了实现这一目标，我们使用mRNA展示和高通量测序技术对〜5×10 ^13个随机的30-mer肽进行了分析，以寻找可以与水溶性全氟芳烃进行半胱氨酸芳基化的多肽。在这个广阔的化学空间中，我们发现了一个含半胱氨酸的序列，其二级速率常数为0.29 M ^–1 s ^–1芳基化。N末端和C末端的截短降低了反应速率，变性剂的添加也降低了反应速率。当反应性肽与酶分选酶A共价融合时，我们在单个半胱氨酸位点观察到了区域特异性芳基化作用，而使酶的活性位点半胱氨酸保持不变。综上所述，这些结果表明，与适当的反应性基团匹配时，确定序列的长多肽可以用于水中半胱氨酸的选择性芳基化。

更新日期：2017-12-28

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