Tyrosinase-Catalyzed Peptide Macrocyclization for mRNA Display
Tyrosinase-Catalyzed Peptide Macrocyclization for mRNA Display
复制标题
用于 mRNA 显示的酪氨酸酶催化肽大环化
DOI：
10.1021/jacs.2c12629
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发表时间：
2023
影响因子：
15
通讯作者：
Bowers, Albert A.
中科院分区：
文献类型：
--
作者：
Fleming, Matthew C.;Bowler, Matthew M.;Park, Rodney;Popov, Konstantin I.;Bowers, Albert A.
大环肽的mRNA展示已被证明是发现蛋白质靶标的高亲和力配体的有力技术。然而，已知只有有限数量的环化化学与mRNA展示相容。酪氨酸酶是一种铜依赖性氧化酶，它将酪氨酸酚氧化成亲电子醌，该亲电子醌容易被半胱氨酸巯基攻击。在这里，我们表明，含有酪氨酸和半胱氨酸的肽在酪氨酸酶处理后迅速环化。环化的表征揭示了它广泛适用于多种大环尺寸和支架。我们将联合收割机酪氨酸酶介导的环化与mRNA展示相结合，以发现新的靶向黑色素瘤相关抗原A4（MAGE-A4）的大环配体。这些大环化合物以纳摩尔IC 50值有效抑制MAGE-A4结合轴。重要的是，大环配体显示出明显的优势，非环化类似物的IC 50值降低了40倍或更多。
mRNA display of macrocyclic peptides has proven itself to be a powerful technique to discover high-affinity ligands for a protein target. However, only a limited number of cyclization chemistries are known to be compatible with mRNA display. Tyrosinase is a copper-dependent oxidase that oxidizes tyrosine phenol to an electrophilico-quinone, which is readily attacked by cysteine thiol. Here we show that peptides containing tyrosine and cysteine are rapidly cyclized upon tyrosinase treatment. Characterization of the cyclization reveals it to be widely applicable to multiple macrocycle sizes and scaffolds. We combine tyrosinase-mediated cyclization with mRNA display to discover new macrocyclic ligands targeting melanoma-associated antigen A4 (MAGE-A4). These macrocycles potently inhibit the MAGE-A4 binding axis with nanomolar IC50values. Importantly, macrocyclic ligands show clear advantage over noncyclized analogues with ∼40-fold or greater decrease in IC50values.