Fusion proteins play an essential role in the biosciences but suffer from several key limitations, including the requirement for N-to-C terminal ligation, incompatibility of constituent domains, incorrect folding, and loss of biological activity. This perspective focuses on chemical and enzymatic approaches for the post-translational generation of well-defined protein–protein conjugates, which overcome some of the limitations faced by traditional fusion techniques. Methods discussed range from chemical modification of nucleophilic canonical amino acid residues to incorporation of unnatural amino acid residues and a range of enzymatic methods, including sortase-mediated ligation. Through summarizing the progress in this rapidly growing field, the key successes and challenges associated with using chemical and enzymatic approaches are highlighted and areas requiring further development are discussed.

中文翻译：

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翻译后蛋白质-蛋白质缀合的化学和酶法


融合蛋白在生物科学中发挥着重要作用，但存在一些关键限制，包括需要 N 至 C 末端连接、组成域不相容、不正确折叠和生物活性丧失。这一观点重点关注用于翻译后生成明确的蛋白质-蛋白质缀合物的化学和酶促方法，这克服了传统融合技术面临的一些限制。讨论的方法范围从亲核规范氨基酸残基的化学修饰到非天然氨基酸残基的掺入以及一系列酶促方法，包括分选酶介导的连接。通过总结这一快速发展领域的进展，强调了与使用化学和酶方法相关的关键成功和挑战，并讨论了需要进一步发展的领域。