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Exploring Structure and Function of Redox Intermediates in [NiFe]‐Hydrogenases by an Advanced Experimental Approach for Solvated, Lyophilized and Crystallized Metalloenzymes
Author(s) -
Lorent Christian,
Pelmenschikov Vladimir,
Frielingsdorf Stefan,
Schoknecht Janna,
Caserta Giorgio,
Yoda Yoshitaka,
Wang Hongxin,
Tamasaku Kenji,
Lenz Oliver,
Cramer Stephen P.,
Horch Marius,
Lauterbach Lars,
Zebger Ingo
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.202100451
Subject(s) - hydrogenase , chemistry , redox , raman spectroscopy , hydride , resonance raman spectroscopy , density functional theory , catalysis , infrared spectroscopy , spectroscopy , reaction intermediate , crystallography , photochemistry , inorganic chemistry , computational chemistry , hydrogen , organic chemistry , physics , quantum mechanics , optics
To study metalloenzymes in detail, we developed a new experimental setup allowing the controlled preparation of catalytic intermediates for characterization by various spectroscopic techniques. The in situ monitoring of redox transitions by infrared spectroscopy in enzyme lyophilizate, crystals, and solution during gas exchange in a wide temperature range can be accomplished as well. Two O 2 ‐tolerant [NiFe]‐hydrogenases were investigated as model systems. First, we utilized our platform to prepare highly concentrated hydrogenase lyophilizate in a paramagnetic state harboring a bridging hydride. This procedure proved beneficial for 57 Fe nuclear resonance vibrational spectroscopy and revealed, in combination with density functional theory calculations, the vibrational fingerprint of this catalytic intermediate. The same in situ IR setup, combined with resonance Raman spectroscopy, provided detailed insights into the redox chemistry of enzyme crystals, underlining the general necessity to complement X‐ray crystallographic data with spectroscopic analyses.