Bietti Antonio L Figueroa, Kauth Alisa-Maite A, Hommel Katrin, Blueggel Mike, Mohr Laurenz, Niemeyer Felix C, Beuck Christine, Bayer Peter, Knauer Shirley K, Jan Ravoo Bart, Schrader Thomas
Institute of Organic Chemistry I, Biosupramolecular Chemistry, University of Duisburg-Essen Universitätsstrasse 7 45141 Essen Germany
Organic Chemistry Institute and Center for Soft Nanoscience, University of Münster Busso-Peus-Straße 10 48149 Münster Germany
RSC Chem Biol. 2025 Sep 5. doi: 10.1039/d5cb00069f.
Light serves as an exceptional stimulus for the precise spatiotemporal regulation of protein activity and protein-protein interactions. Here, we introduce a light-responsive supramolecular ligand system designed to modulate Taspase 1, a protease critical for embryogenesis and implicated in tumor progression. Our approach utilizes photoswitchable divalent molecular tweezers engineered to target lysine-rich regions within the Taspase 1 loop. By incorporating arylazopyrazole (AAP) photoswitches, we achieve dynamic and reversible control of ligand binding. These photoswitches exhibit high photostationary states, excellent reversibility, and prolonged thermal stability of the isomer, ensuring reliable switching without photodegradation. The tweezer distance varies between and isomers, enabling tunable binding interactions. Through a combination of surface plasmon resonance, enzymatic cleavage assays, and molecular dynamics simulations, we demonstrate that these ligands bind Taspase 1 with low micromolar affinity and effectively inhibit its proteolytic activity. While isomerization did not significantly affect the inhibition of protein-protein interaction, the -isomers of larger tweezers exhibited powerful enzyme inhibition, likely due to their ability to bridge lysines flanking the active site. This photoswitchable tweezer system provides a versatile tool for light-controlled modulation of protein function, offering new opportunities for selectively targeting lysine-rich proteins in dynamic biological environments.
光作为一种特殊的刺激因素，可对蛋白质活性和蛋白质 - 蛋白质相互作用进行精确的时空调节。在此，我们介绍一种光响应超分子配体系统，该系统旨在调节Taspase 1，Taspase 1是一种对胚胎发育至关重要且与肿瘤进展相关的蛋白酶。我们的方法利用了经过工程改造的可光开关二价分子镊子，以靶向Taspase 1环内富含赖氨酸的区域。通过引入芳基偶氮吡唑（AAP）光开关，我们实现了配体结合的动态和可逆控制。这些光开关表现出高光稳态、出色的可逆性以及异构体的延长热稳定性，确保了可靠的切换且无光降解。镊子距离在异构体之间变化，从而实现可调节的结合相互作用。通过表面等离子体共振、酶切测定和分子动力学模拟相结合，我们证明这些配体以低微摩尔亲和力结合Taspase 1并有效抑制其蛋白水解活性。虽然异构化对蛋白质 - 蛋白质相互作用的抑制没有显著影响，但较大镊子的 - 异构体表现出强大的酶抑制作用，这可能是由于它们能够桥接活性位点两侧的赖氨酸。这种可光开关的镊子系统为蛋白质功能的光控调节提供了一种通用工具，为在动态生物环境中选择性靶向富含赖氨酸的蛋白质提供了新机会。
