Exploring Structure and Function of Redox Intermediates in [NiFe]-Hydrogenases by an Advanced Experimental Approach for Solvated, Lyophilized and Crystallized Metalloenzymes.

ABSTRACT: 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 O₂ -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

SUBMITTER: Lorent C 

PROVIDER: S-EPMC8360142 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature