Three-megadalton complex of methanogenic electron-bifurcating and CO2-fixing enzymes

• Published in: Science (American Association for the Advancement of Science) Vol. 373; no. 6559; pp. 1151 - 1156
• Main Authors: Watanabe, Tomohiro, Pfeil-Gardiner, Olivia, Kahnt, Jörg, Koch, Jürgen, Shima, Seigo, Murphy, Bonnie J
• Format: Journal Article
• Language: English
• Published: Washington The American Association for the Advancement of Science 03.09.2021
• Subjects: Bifurcations; Carbon dioxide; Cofactors; Dehydrogenase; Dehydrogenases; Electron microscopy; Enzymes; Ferredoxin; Flavin; Formate dehydrogenase; Heterodisulfide reductase; Hydrogenase; Methanogenesis; Methanogenic bacteria; Oxidation; Reductases; Reduction; Substrates; Sulfur
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.abg5550

Methanogenesis megacomplexAn important first step in methanogenesis is the conversion of carbon dioxide to a reduced one-carbon formyl unit that is a substrate for downstream steps. This reaction is catalyzed by a complex of enzymes, including components for oxidizing hydrogen or formate and splitting two electrons along different energetic paths. Watanabe et al. carefully purified and prepared anaerobic cryo–electron microscopy samples of the enzyme complex responsible, resulting in a three-megadalton hexameric structure at 3- to 3.5-ångström resolution. The arrangement of iron–sulfur cofactors provides an explanation for how electron bifurcation is coupled to large protein motions, which are expected from the multiple conformational states present. —MAFThe first reaction of the methanogenic pathway from carbon dioxide (CO2) is the reduction and condensation of CO2 to formyl-methanofuran, catalyzed by formyl-methanofuran dehydrogenase (Fmd). Strongly reducing electrons for this reaction are generated by heterodisulfide reductase (Hdr) in complex with hydrogenase or formate dehydrogenase (Fdh) using a flavin-based electron-bifurcation mechanism. Here, we report enzymological and structural characterizations of Fdh-Hdr-Fmd complexes from Methanospirillum hungatei. The complexes catalyze this reaction using electrons from formate and the reduced form of the electron carrier F420. Conformational changes in HdrA mediate electron bifurcation, and polyferredoxin FmdF directly transfers electrons to the CO2 reduction site, as evidenced by methanofuran-dependent flavin-based electron bifurcation even without free ferredoxin, a diffusible electron carrier between Hdr and Fmd. Conservation of Hdr and Fmd structures suggests that this complex is common among hydrogenotrophic methanogens.