title: Preparation of [Fe]-hydrogenase from methanogenic archaea.
abstract: [Fe]-hydrogenase is one of the three types of hydrogenases. This enzyme is found in many hydrogenotrophic methanogenic archaea and catalyzes the reversible hydride transfer from H(2) to methenyl-H(4)MPT(+) in methanogenesis from H(2) and CO(2). The enzyme harbors a unique iron-guanylyl pyridinol (FeGP) cofactor as a prosthetic group. Here, we describe the purification of [Fe]-hydrogenase from Methanothermobacter marburgensis, the isolation of the FeGP cofactor from the native holoenzyme, and the reconstitution of [Fe]-hydrogenase from the isolated FeGP cofactor and the heterologously produced apoenzyme.
authors.affiliation: Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
authors.forename: Seigo
authors.initials: S
authors.lastname: Shima
authors.forename: Michael
authors.initials: M
authors.lastname: Schick
authors.forename: Haruka
authors.initials: H
authors.lastname: Tamura
mesh_terms: D001105:Archaea / Q000201:enzymology*; D019843:Archaeal Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D006864:Hydrogenase / Q000302:isolation & purification* / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D008697:Methane / Q000378:metabolism*; D017013:Methanobacteriaceae / Q000201:enzymology
chemical_list: D019843:Archaeal Proteins; D007506:Iron-Sulfur Proteins; C118830:iron hydrogenase; D006864:Hydrogenase; D008697:Methane