Sondermann Philipp, Diercks Christian S, Rong Cynthia, Schultz Peter G
Department of Chemistry and Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037.
Proc Natl Acad Sci U S A. 2025 Apr;122(13):e2426745122. doi: 10.1073/pnas.2426745122. Epub 2025 Mar 24.
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 α-Synuclein, a presynaptic 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 α-Synuclein. The protease's selectivity is further demonstrated in human cells where near complete degradation of overexpressed human α-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.
人们对靶向降解与人类疾病相关的蛋白质有着浓厚的兴趣。为此目的使用序列特异性蛋白酶受到严重限制，因为在设计产生高选择性蛋白酶所需的众多酶 - 底物相互作用时存在困难，同时还要保持催化活性。在此，我们报告了一种策略，用于进化一种蛋白酶，以实现与帕金森病密切相关的突触前蛋白α-突触核蛋白的程序性降解。我们基于结构的进化研究使用了肉毒杆菌神经毒素中的蛋白酶，并展示了其特异性从天然底物SNAP25逐步转变为对α-突触核蛋白的选择性降解。该蛋白酶的选择性在人类细胞中进一步得到证明，在那里观察到过表达的人类α-突触核蛋白几乎完全降解，而对细胞增殖没有显著影响。这种逐步策略可能作为一种通用方法，用于进化针对失调蛋白的高选择性蛋白酶。
