Targeted degradation of α- Synuclein using an evolved botulinum toxin protease

• 成果类型: Article
• 署名作者: Sondermann, Philipp; Diercks, Christian S.; Rong, Cynthia; Schultz, Peter G.
• 署名单位: Scripps Research Institute; Scripps Research Institute
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-15292
• DOI: 10.1073/pnas.2426745122
• 发表日期: 2025-04-01
• 关键词: neurotoxin; dna; mutations; vector

摘要

There is considerable interest in the targeted degradation of proteins implicated in human disease. The use of sequence-specific proteases for this purpose is severely limited by the difficulty in engineering the numerous enzyme-substrate interactions required to yield highly selective proteases while maintaining catalytic activity. Herein, we report a strategy to evolve a protease for the programmed degradation of alpha- Synuclein, a pre-synaptic protein closely linked to Parkinson's disease. Our structure-guided evolution campaign uses the protease from botulinum neurotoxin and showcases the stepwise change of specificity from its native substrate SNAP25 to the selective degradation of alpha- Synuclein. The protease's selectivity is further demonstrated in human cells where near complete degradation of overexpressed human alpha- Synuclein is observed with no significant effects on cell proliferation. This stepwise strategy may serve as a general approach to evolve highly selective proteases targeting dysregulated proteins.