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A Genetically Encoded Two‐Dimensional Infrared Probe for Enzyme Active‐Site Dynamics
Author(s) -
Wang Li,
Zhang Jia,
Han MingJie,
Zhang Lu,
Chen Chao,
Huang Aiping,
Xie Ruipei,
Wang Guosheng,
Zhu Jiangrui,
Wang Yuchuan,
Liu Xiaohong,
Zhuang Wei,
Li Yunliang,
Wang Jiangyun
Publication year - 2021
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202016880
Subject(s) - active site , chemistry , enzyme , protein dynamics , biophysics , photochemistry , biochemistry , protein structure , biology
While two‐dimensional infrared (2D‐IR) spectroscopy is uniquely suitable for monitoring femtosecond (fs) to picosecond (ps) water dynamics around static protein structures, its utility for probing enzyme active‐site dynamics is limited due to the lack of site‐specific 2D‐IR probes. We demonstrate the genetic incorporation of a novel 2D‐IR probe, m ‐azido‐ L ‐tyrosine (N3Y) in the active‐site of DddK, an iron‐dependent enzyme that catalyzes the conversion of dimethylsulfoniopropionate to dimethylsulphide. Our results show that both the oxidation of active‐site iron to Fe III , and the addition of denaturation reagents, result in significant decrease in enzyme activity and active‐site water motion confinement. As tyrosine residues play important roles, including as general acids and bases, and electron transfer agents in many key enzymes, the genetically encoded 2D‐IR probe N3Y should be broadly applicable to investigate how the enzyme active‐site motions at the fs–ps time scale direct reaction pathways to facilitating specific chemical reactions.