Phage-assisted evolution of botulinum neurotoxin proteases with reprogrammed specificity

doi:10.1126/science.abf5972

GSTDTAP > 气候变化

DOI	10.1126/science.abf5972
	Phage-assisted evolution of botulinum neurotoxin proteases with reprogrammed specificity
	Travis R. Blum; Hao Liu; Michael S. Packer; Xiaozhe Xiong; Pyung-Gang Lee; Sicai Zhang; Michelle Richter; George Minasov; Karla J. F. Satchell; Min Dong; David R. Liu
	2021-02-19
发表期刊	Science
出版年	2021
英文摘要	Proteases that cleave protein targets at specific sequences control many biological functions. The ability to reprogram proteases to cleave new sequences of our choosing would enable new therapeutic and biotechnological applications. Blum et al. report a laboratory evolution method to rapidly evolve proteases that cut new protein sequences and lose their ability to cut nontarget sequences (see the Perspective by Stenmark). Using this method, they evolved botulinum neurotoxin proteases, an important class of enzymes used in patients, to selectively cleave new targets, including a protein unrelated to those natively cleaved by these proteases. This work establishes a powerful approach to generate proteases with tailor-made specificities. Science , this issue p. [803][1]; see also p. [782][2] Although bespoke, sequence-specific proteases have the potential to advance biotechnology and medicine, generation of proteases with tailor-made cleavage specificities remains a major challenge. We developed a phage-assisted protease evolution system with simultaneous positive and negative selection and applied it to three botulinum neurotoxin (BoNT) light-chain proteases. We evolved BoNT/X protease into separate variants that preferentially cleave vesicle-associated membrane protein 4 (VAMP4) and Ykt6, evolved BoNT/F protease to selectively cleave the non-native substrate VAMP7, and evolved BoNT/E protease to cleave phosphatase and tensin homolog (PTEN) but not any natural BoNT protease substrate in neurons. The evolved proteases display large changes in specificity (218- to >11,000,000-fold) and can retain their ability to form holotoxins that self-deliver into primary neurons. These findings establish a versatile platform for reprogramming proteases to selectively cleave new targets of therapeutic interest. [1]: /lookup/doi/10.1126/science.abf5972 [2]: /lookup/doi/10.1126/science.abg3535
领域	气候变化 ; 资源环境
URL	查看原文
引用统计
文献类型	期刊论文
条目标识符	http://119.78.100.173/C666/handle/2XK7JSWQ/315724
专题	气候变化资源环境科学
推荐引用方式 GB/T 7714	Travis R. Blum,Hao Liu,Michael S. Packer,et al. Phage-assisted evolution of botulinum neurotoxin proteases with reprogrammed specificity[J]. Science,2021.
APA	Travis R. Blum.,Hao Liu.,Michael S. Packer.,Xiaozhe Xiong.,Pyung-Gang Lee.,...&David R. Liu.(2021).Phage-assisted evolution of botulinum neurotoxin proteases with reprogrammed specificity.Science.
MLA	Travis R. Blum,et al."Phage-assisted evolution of botulinum neurotoxin proteases with reprogrammed specificity".Science (2021).