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Literature summary for 1.14.13.25 extracted from

  • Srinivas, V.; Banerjee, R.; Lebrette, H.; Jones, J.C.; Aurelius, O.; Kim, I.S.; Pham, C.C.; Gul, S.; Sutherlin, K.D.; Bhowmick, A.; John, J.; Bozkurt, E.; Fransson, T.; Aller, P.; Butryn, A.; Bogacz, I.; Simon, P.; Keable, S.; Britz, A.; Tono, K.; Kim, K.S.; Park, S.Y.; Lee, S.J.; Park, J.; Alonso-Mori, R.; Fu, F.u.l.
    High-resolution XFEL structure of the soluble methane monooxygenase hydroxylase complex with its regulatory component at ambient temperature in two oxidation states (2020), J. Am. Chem. Soc., 142, 14249-14266 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
recombinant enzyme expression in Escherichia coli strain BL21(DE3) Methylosinus trichosporium

Crystallization (Commentary)

Crystallization (Comment) Organism
sMMOH:MMOB complex in fully oxidized and fully reduced states, sitting drop vapor diffusion method, mixing of 0.0225 ml of protein solution containing 0.053 mM sMMOH and 0.106 mM MMOB in 25 mM MOPS, pH 7.0, with 0.0075 ml of reservoir solution containing 100 mM HEPES/MOPS, pH 7.5, 30 mM NaI, 30 mM NaBr, 30 mM NaF, 20% v/v glycerol, 10% w/v PEG 4000, and 10 mM FeCl3, equilibration against 0.5 ml reservoir solution, 3 days at room temperature, X-ray diffraction structure determination and analysis at 1.95-2.9 A resolution. Microcrystals of the sMMOH:MMOB complex from Methylosinus trichosporium OB3b are 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. Structural reorganization analysis. The position of Glu243 relative to Fe2 is shifted to the bridging position. Detailed method overview Methylosinus trichosporium

Metals/Ions

Metals/Ions Comment Organism Structure
Fe the enzyme uses a nonheme, oxygen-bridged dinuclear iron cluster in the active site Methylosinus trichosporium

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
methane + NADH + H+ + O2 Methylosinus trichosporium
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methanol + NAD+ + H2O
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?

Organism

Organism UniProt Comment Textmining
Methylosinus trichosporium A0A2D2D5X0 AND A0A2D2D0T8 AND Q53563 AND A0A2D2D0X7 MMOH, MMOR, MMOB, and MMOD
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methane + NADH + H+ + O2
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Methylosinus trichosporium methanol + NAD+ + H2O
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?

Subunits

Subunits Comment Organism
More the sMMO enzyme consists of three protein components: a 245 kDa (alphabetagamma)2 hydroxylase (MMOH), a 37 kDa FAD and Fe2S2 cluster-containing reductase (MMOR), and a 15 kDa regulatory protein (MMOB). The active site is buried deep within sMMOH and contains an oxygen-bridged dinuclear FeIII cluster in which the irons are bridged by two hydroxo moieties and a carboxylate from Glu144. After reduction, the diiron cluster functions to activate O2 and insert an oxygen atom into a highly stable (105 kcal/mol bond dissociation energy) C-H bond of methane. Although chemically reduced sMMOH can carry out the oxygenation chemistry alone, the reaction only proceeds at a physiologically relevant rate when sMMOH is complexed with MMOB. Reduction of the diiron cluster causes a shift in the position of Glu243, a monodentate ligand to Fe2 in the diferric cluster. In the shifted position, Glu243 bridges Fe1 and Fe2 via one of its carboxylate oxygens, thereby directly displacing one of the bridging solvents. The second bridging solvent bond is weakened, which presumably allows facile displacement by O2 to begin the oxygen activation process Methylosinus trichosporium
trimer 1 * 245000, alpha-subunitMMOH, + 1 * 37000, beta-subunit MMOR, + 1 * 15000, gamma-subunit MMOB Methylosinus trichosporium

Synonyms

Synonyms Comment Organism
sMMO
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Methylosinus trichosporium
soluble methane monooxygenase
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Methylosinus trichosporium

Cofactor

Cofactor Comment Organism Structure
FAD
-
Methylosinus trichosporium
NADH
-
Methylosinus trichosporium

General Information

General Information Comment Organism
additional information enzyme sMMO structural reorganization through subunits MMOH and MMOB including the dinuclear iron cluster, detailed overview Methylosinus trichosporium
physiological function 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 O2 activation and substrate hydroxylation. 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. Although chemically reduced sMMOH can carry out the oxygenation chemistry alone, the reaction only proceeds at a physiologically relevant rate when sMMOH is complexed with MMOB. Many regulatory functions of MMOB have been discovered but its most important effects are to decrease the redox potential of the diiron cluster by 132 mV, accelerate O2 binding by 1000fold, increase the turnover number 150fold, and tune sMMOH to selectively bind and oxygenate methane over other more easily oxidized hydrocarbons Methylosinus trichosporium