High-Resolution XFEL Structure of the Soluble Methane Monooxygenase Hydroxylase Complex with its Regulatory Component at Ambient Temperature in Two Oxidation States
| Autor: | Hugo Lebrette, Kensuke Tono, Kyle D. Sutherlin, Nicholas K. Sauter, Sang Jae Lee, Cindy C. Pham, Vittal K. Yachandra, Junko Yano, A. Butryn, Pierre Aller, Thomas Fransson, Allen M. Orville, Uwe Bergmann, Martin Högbom, Franklin D. Fuller, Sang-Youn Park, Oskar Aurelius, Alexander Britz, Aaron S. Brewster, Sheraz Gul, Isabel Bogacz, Kyung Sook Kim, Stephen Keable, Juliane John, Vivek Srinivas, In-Sik Kim, Alexander Batyuk, Philipp S. Simon, John D. Lipscomb, Roberto Alonso-Mori, Jason C. Jones, Asmit Bhowmick, Esra Bozkurt, Jae Hyun Park, Rahul Banerjee, Jan Kern |
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| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular Methane monooxygenase Crystal structure 010402 general chemistry Photochemistry 01 natural sciences Biochemistry Catalysis Methane Article Hydroxylation Metal chemistry.chemical_compound Colloid and Surface Chemistry Models biology X-Rays Temperature Active site Substrate (chemistry) Molecular General Chemistry Methylosinus trichosporium 0104 chemical sciences chemistry Solubility visual_art Anaerobic oxidation of methane Chemical Sciences biology.protein visual_art.visual_art_medium Oxygenases Oxidation-Reduction |
| Zdroj: | Journal of the American Chemical Society, vol 142, iss 33 J Am Chem Soc Journal of the American Chemical Society |
| Popis: | Soluble methane monooxygenase (sMMO) is a multicomponent metalloenzyme that catalyzes the conversion of methane to methanol at ambient temperature using a nonheme, oxygen-bridged dinuclear iron cluster in the active site. Structural changes in the hydroxylase component (sMMOH) containing the diiron cluster caused by complex formation with a regulatory component (MMOB) and by iron reduction are important for the regulation of O(2) activation and substrate hydroxylation. Structural studies of metalloenzymes using traditional synchrotron-based X-ray crystallography are often complicated by partial X-ray-induced photoreduction of the metal center, thereby obviating determination of the structure of the enzyme in pure oxidation states. Here microcrystals of the sMMOH:MMOB complex from Methylosinus trichosporium OB3b were serially exposed to X-ray free electron laser (XFEL) pulses, where the [35 fs duration of exposure of an individual crystal yields diffraction data before photoreduction-induced structural changes can manifest. Merging diffraction patterns obtained from thousands of crystals generates radiation damage free, 1.95 Å resolution crystal structures for the fully oxidized and fully reduced states of the sMMOH:MMOB complex for the first time. The results provide new insight into the manner by which the diiron cluster and the active site environment are reorganized by the regulatory protein component in order to enhance the steps of oxygen activation and methane oxidation. This study also emphasizes the value of XFEL and serial femtosecond crystallography (SFX) methods for investigating the structures of metalloenzymes with radiation sensitive metal active sites. |
| Databáze: | OpenAIRE |
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