6GM7
[FeFe]-hydrogenase HydA1 from Chlamydomonas reinhardtii,variant E144A
Summary for 6GM7
| Entry DOI | 10.2210/pdb6gm7/pdb |
| Related | 3LX4 4R0V |
| Descriptor | Fe-hydrogenase, IRON/SULFUR CLUSTER, CHLORIDE ION, ... (4 entities in total) |
| Functional Keywords | hydrogenase, h-cluster, apo-form, oxidoreductase |
| Biological source | Chlamydomonas reinhardtii (Chlamydomonas smithii) |
| Total number of polymer chains | 1 |
| Total formula weight | 49206.78 |
| Authors | Duan, J.,Engelbrecht, V.,Esselborn, J.,Hofmann, E.,Winkler, M.,Happe, T. (deposition date: 2018-05-24, release date: 2018-11-07, Last modification date: 2024-05-15) |
| Primary citation | Duan, J.,Senger, M.,Esselborn, J.,Engelbrecht, V.,Wittkamp, F.,Apfel, U.P.,Hofmann, E.,Stripp, S.T.,Happe, T.,Winkler, M. Crystallographic and spectroscopic assignment of the proton transfer pathway in [FeFe]-hydrogenases. Nat Commun, 9:4726-4726, 2018 Cited by PubMed Abstract: The unmatched catalytic turnover rates of [FeFe]-hydrogenases require an exceptionally efficient proton-transfer (PT) pathway to shuttle protons as substrates or products between bulk water and catalytic center. For clostridial [FeFe]-hydrogenase CpI such a pathway has been proposed and analyzed, but mainly on a theoretical basis. Here, eleven enzyme variants of two different [FeFe]-hydrogenases (CpI and HydA1) with substitutions in the presumptive PT-pathway are examined kinetically, spectroscopically, and crystallographically to provide solid experimental proof for its role in hydrogen-turnover. Targeting key residues of the PT-pathway by site directed mutagenesis significantly alters the pH-activity profile of these variants and in presence of H their cofactor is trapped in an intermediate state indicative of precluded proton-transfer. Furthermore, crystal structures coherently explain the individual levels of residual activity, demonstrating e.g. how trapped HO molecules rescue the interrupted PT-pathway. These features provide conclusive evidence that the targeted positions are indeed vital for catalytic proton-transfer. PubMed: 30413719DOI: 10.1038/s41467-018-07140-x PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.14 Å) |
Structure validation
Download full validation report
