8QM3

formaldehyde-inhibited [FeFe]-hydrogenase I from Clostridium pasteurianum (CpI)

Summary for 8QM3

• Entry DOI: 10.2210/pdb8qm3/pdb
• Related: 4XDC
• Descriptor: Iron hydrogenase 1, dicarbonyl[bis(cyanide-kappaC)]-mu-(iminodimethanethiolatato-1kappaS:2kappaS)-mu-(oxomethylidene)diiron(2+), IRON/SULFUR CLUSTER, ... (9 entities in total)
• Functional Keywords: [fefe]-hydrogenase, formaldehyde-inhibited states, oxidoreductase
• Biological source: Clostridium pasteurianum
• Total number of polymer chains: 2
• Total formula weight: 135135.82

Authors

Duan, J.,Hofmann, E. (deposition date: 2023-09-21, release date: 2023-11-29, Last modification date: 2024-11-20)

Primary citation

Duan, J.,Veliju, A.,Lampret, O.,Liu, L.,Yadav, S.,Apfel, U.P.,Armstrong, F.A.,Hemschemeier, A.,Hofmann, E.
Insights into the Molecular Mechanism of Formaldehyde Inhibition of [FeFe]-Hydrogenases.
J.Am.Chem.Soc., 145:26068-26074, 2023

PubMed Abstract: [FeFe]-hydrogenases are efficient H converting biocatalysts that are inhibited by formaldehyde (HCHO). The molecular mechanism of this inhibition has so far not been experimentally solved. Here, we obtained high-resolution crystal structures of the HCHO-treated [FeFe]-hydrogenase CpI from , showing HCHO reacts with the secondary amine base of the catalytic cofactor and the cysteine C299 of the proton transfer pathway which both are very important for catalytic turnover. Kinetic assays via protein film electrochemistry show the CpI variant C299D is significantly less inhibited by HCHO, corroborating the structural results. By combining our data from protein crystallography, site-directed mutagenesis and protein film electrochemistry, a reaction mechanism involving the cofactor's amine base, the thiol group of C299 and HCHO can be deduced. In addition to the specific case of [FeFe]-hydrogenases, our study provides additional insights into the reactions between HCHO and protein molecules.

PubMed: 37983562
DOI: 10.1021/jacs.3c07800

Experimental method

X-RAY DIFFRACTION (1.53 Å)