A TeZla micromixer for interrogating the early and broad folding landscape of G-quadruplex via multistage velocity descending

• 成果类型: Article
• 署名作者: Li, Zheyu; Hu, Rui; Li, Tao; Zhu, Jiang; You, Huijuan; Li, Yiwei; Liu, Bi-Feng; Li, Conggang; Li, Ying; Yang, Yunhuang
• 署名单位: Huazhong University of Science & Technology; Chinese Academy of Sciences; Chinese Academy of Sciences; University of Chinese Academy of Sciences, CAS; Huazhong University of Science & Technology; Huazhong University of Science & Technology
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-13085
• DOI: 10.1073/pnas.2315401121
• 发表日期: 2024-01-23
• 关键词: stranded g-quadruplexes; microfluidic mixer; loop length; dna; kinetics; fluorescence; STABILITY; pathways; DYNAMICS; mechanism

摘要

Biomacromolecular folding kinetics involves fast folding events and broad timescales. Current techniques face limitations in either the required time resolution or the observation window. In this study, we developed the TeZla micromixer, integrating Tesla and Zigzag microstructures with a multistage velocity descending strategy. TeZla achieves a significant short mixing dead time (40 mu s) and a wide time window covering four orders of magnitude (up to 300 ms). Using this unique micromixer, we explored the folding landscape of c-Myc G4 and its noncanonical-G4 derivatives with different loop lengths or G-vacancy sites. Our findings revealed that c-Myc can bypass folding intermediates and directly adopt a G4 structure in the cation-deficient buffer. Moreover, we found that the loop length and specific G-vacancy site could affect the folding pathway and significantly slow down the folding rates. These results were also cross-validated with real-time NMR and circular dichroism. In conclusion, TeZla represents a versatile tool for studying biomolecular folding kinetics, and our findings may ultimately contribute to the design of drugs targeting G4 structures.