Abstract

Making peptide bonds is tightly controlled by genetic code and machinery which includes cofactors, ATP, and RNAs. In this regard, the stand-alone and genetic-code-independent peptide ligases constitute a new family of renegade peptide-bond makers. A prime example is butelase-1, an Asn/Asp(Asx)-specific ligase that structurally belongs to the asparaginyl endopeptidase family. Butelase-1 specifically recognizes a C-terminal Asx-containing tripeptide motif, Asn/Asp-Xaa-Yaa (Xaa and Yaa are any amino acids), to form a site-specific Asn-Xaa peptide bond either intramolecularly as cyclic proteins or intermolecularly as modified proteins. Our work in the past five years has validated that butelase-1 is a potent and versatile ligase. Here we review the advances in ligases, with a focus on butelase-1, and their applications in engineering bioactive peptides and precision protein modifications, antibody-drug conjugates, and live-cell labeling.

中文翻译：

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肽天冬酰胺基连接酶-叛逆性肽键形成者

摘要

肽键的形成受到遗传密码和机制（包括辅因子，ATP和RNA）的严格控制。在这方面，独立的和不依赖遗传密码的肽连接酶构成了新的反叛肽键制造者家族。一个主要的例子是butelase-1，这是一种Asn / Asp（Asx）特异性连接酶，结构上属于天冬酰胺基内肽酶家族。Butelase-1特异性识别C端含Asx的三肽基序Asn / Asp-Xaa-Yaa（Xaa和Yaa是任何氨基酸），以分子内或环状分子的形式形成位点特异性Asn-Xaa肽键作为修饰的蛋白质。在过去的五年中，我们的工作已经验证了butelase-1是一种有效且用途广泛的连接酶。在这里，我们重点介绍连接酶的研究进展，重点是butelase-1，