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Real-Time Observation of Protein Transport across Membranes by Femtosecond Sum Frequency Generation Vibrational Spectroscopy
Junjun Tan1,2, Chuanzhao Li1,2, Jiahui Zhang1,2, Shuji Ye1,2
1.Hefei National Laboratory for Physical Sciences at the Microscale, and Department of Chemical Physics, University of Science and Technology of China, Hefei 230026, China;2.Synergetic Innovation Center of Quantum Information & Quantum Physics, University of Science and Technology of China, Hefei 230026, China
Abstract:
Characterization of conformation kinetics of proteins at the interfaces is crucial for understanding the biomolecular functions and the mechanisms of interfacial biological action. But it requires to capture the dynamic structures of proteins at the interfaces with sufficient structural and temporal resolutions. Here, we demonstrate that a femtosecond sum frequency generation vibrational spectroscopy (SFG-VS) system developed by our group provides a powerful tool for monitoring the real-time peptide transport across the membranes with time resolution of less than one second. By probing the real-time SFG signals in the amide I and amide A bands as WALP23 interacts with DMPG lipid bilayer, it is found that WALP23 is initially absorbed at the gel-phase DMPG bilayer with a random coil structure. The absorption of WALP23 on the surface leads to the surface charge reversal and thus changes the orientation of membrane-bound water. As the DMPG bilayer changes from gel phase into fluid phase, WALP23 inserts into the fluid-phase bilayer with its N-terminal end moving across the membrane, which causes the membrane dehydration and the transition of WALP23 conformation from random coil to mixed helix/loop structure and then to pure α-helical structure. The established system is ready to be employed in characterizing other interfacial fast processes, which will be certainly helpful for providing a clear physical picture of the interfacial phenomena.
Key words:  Femtosecond sum frequency generation  Peptide transport across membrane  Real time  Kinetic conformation
FundProject:This work was supported by the National Natural Science Foundation of China (No.21473177, No.21633007), the National Key Research and Development Program of China (No.2017YFA0303500 and No.2018YFA0208700), Fundamental Research Funds for the Central Universities (No.WK2340000064), Anhui Initiative in Quantum Information Technologies (No.AHY090000).
蛋白质跨膜传输的飞秒和频振动光谱快速实时监测
谈军军1,2, 李传召1,2, 张佳慧1,2, 叶树集1,2
1.中国科学技术大学合肥微尺度物质科学国家研究中心, 化学物理系, 合肥 230026;2.中国科学技术大学量子信息与量子科技前沿协同创新中心, 能源材料化学协同创新中心, 合肥 230026
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DOI:10.1063/1674-0068/31/cjcp1805128
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