Heterodisulfide reductase / [NiFe]-hydrogenase complex from Methanothermococcus thermolithotrophicus soaked with heterodisulfide for 3.5 minutes

Methanogenic heterodisulfide reductase (HdrABC-MvhAGD) uses two noncubane [4Fe-4S] clusters for reduction.

This subunit contains two pentacoordinated non cubane [4Fe-4S] clusters

Methyl-viologen reducing hydrogenase, subunit D

Methyl-viologen reducing hydrogenase, subunit G

The C-terminus has been already post-translationally cleaved
Methyl-viologen reducing hydrogenase, subunit A

Methanothermococcus thermolithotrophicus DSM 2095

Black thick plate square shape of about 0.2-0.4 mm

All crystallization was performed in an anaerobic chamber (95% N2/5% H2) with anoxic solution using the sitting drop method (96-well 2-drop MRC Crystallization Plates in polystyrene, Molecular Dimensions, Suffolk, UK). The crystallization reservoir contained 100 mM Tris/HCl, pH 8.5, 3% (v/v) DMSO, 30% (w/v) polyethylene glycol 4000, and 200 mM Na acetate trihydrate. Crystallization drop contained 1 ul HdrABC-MvhAGD at 25 mg/ml premixed with 2 mM FAD and 1 ul of precipitant. The crystals appeared after 1-2 weeks in this condition. The cryo-cool trapping experiment was performed as following: crystals were soaked for 3min30sec in the crystallization solution supplemented with 66 mM CoM-S-S-CoB. The crystal was cryo-protected by a soak in the crystallization solution supplemented by 30% glycerol and 10 mM CoM-S-S-CoB.

FAD dependent oxidoreductase, CoB/CoM heterodisulfide reductase, subunit B (E.C.1.8.98.1), CoB/CoM heterodisulfide reductase, subunit C, Methyl-viologen-reducing hydrogenase delta subunit, Hydrogen dehydrogenase (E.C.1.12.1.2)
Heterodisulfide reductase, [NiFe]-hydrogenase, FeS cluster, ferredoxin, CCG motif, methanogenesis, flavoprotein, flavin-based electron bifurcation, [2Fe-2S] cluster, macromolecular complex, anaerobic, thioredoxin, metabolism, OXIDOREDUCTASE