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1.14.13.25: methane monooxygenase (soluble)

This is an abbreviated version!
For detailed information about methane monooxygenase (soluble), go to the full flat file.

Word Map on EC 1.14.13.25

Reaction

methane
+
NAD(P)H
 +
H+
 +
O2
=
methanol
 +
NAD(P)+
 +
H2O

Synonyms

chcA, cytoplasmic methane monooxygenase, methane hydroxylase, methane mono-oxygenase, methane monooxygenase, methane monooxygenase hydroxylase, MmMmoC, MMO, MMO Bath, MMOB, MmoC, MMOH, MMOR, oxygenase, methane mono-, particulate methane monooxygenase, pMMO, sMMO, soluble methane monooxygenase, soluble methane monooxygenase hydroxylase

ECTree

     1 Oxidoreductases
         1.14 Acting on paired donors, with incorporation or reduction of molecular oxygen
             1.14.13 With NADH or NADPH as one donor, and incorporation of one atom of oxygen into the other donor
                1.14.13.25 methane monooxygenase (soluble)

Crystallization

Crystallization on EC 1.14.13.25 - methane monooxygenase (soluble)

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis
-
enzyme sMMO crystal structure analysis, PDB ID 1MTY
P22869; P18798; P11987; P18797; P22868; P22867
sitting drop vapor diffusion method, using 0.1 M MES (pH 6.5) and 15% PEG 20000 (w/v)
X-ray crystal structure of the apo, Mn(II)-soaked, and Co(II)-grown hydroxylase component of methane monooxygenase, determined at 2.3 A or greater resolution, reveal that the presence of metal ions is essential for the proper folding of helices E, F, and H of the alpha-subunit
-
purified recombinant MMOH-MMOD complex, hanging drop vapor diffusion method, mixing of 10 mg/ml subunits protein solutions containing 30 mM HEPES, pH 7.5, 100 mM NaCl, and 1 mM TCEP in a 1:2 molar ratio, and mixing with the crystallization solution containing with a mixture of 10% w/v PEG 8000, 20% v/vethylene glycol, 0.02 M 1,6-hexanediol, 0.02 M 1-butanol, 0.02 M 1,2-propanediol, 0.02 M 2-propanol, 0.02 M 1,4-butanediol, 0.02 M 1,3-propanediol, 0.89 M 1,3-butanediol, and 0.1 M MES/imidazole buffer, pH 6.5, one week at room temperature, X-ray diffraction structure determination and analysis at 2.6 A resolution, molecular replacement using Methylosinus trichosporium strain OB3b MMOH as the search model (PDB ID 1MHY), and modeling
-
analysis of the cyrstal structure of the sMMOH:5FWMMOB complex (PDB ID 7M8Q) showing the interface region containing 5FW76 and 5FW77, and of the structure of the sMMOH:BTFA-K15C-5FW-MMOB complex (PDB ID 7M8R) interface region showing the relative position of the BTFA and 5FW 19F-labels
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7
crystallization of wild-type and mutant enzyme complexes by sitting drop vapour diffusion method, for the sMMOH:DBL2 and sMMOH:H5A proteins, 0.0015 ml of protein solution containing 0.06 mM sMMOH and 0.12 mM of either DBL2 or H5A in 100 mM MOPS buffer, pH 7.0, with 0.0015 ml cyrstallization solution containing 20% PEG 3350 and 0.2 M Na2HPO4, pH 8.8, equilibration against 0.5 ml of crystallization solution, 2-3 days at room temperature. For the sMMOH:DBL1 and sMMOH:H33A proteins, 0.0015 ml of protein solution containing 0.06 mM sMMOH and 0.12 mM of either DBL1 or H33A in 100 mM MOPS buffer, pH 7.0, with 0.0015 ml cyrstallization solution containing 21% PEG 3350 and 0.2 M Na2HPO4, pH 6.6, equilibration against 0.5 ml of crystallization solution, 2-3 days at room temperature, X-ray diffraction structure determination and analysis at 1.82-2.40 A resolution, structure modeling
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7
diferric and diferrous states of both sMMOH and the sMMOH:MMOB complexes, X-ray diffraction structure determination and analysis at 1.52-2.35 A resolution, the structures are analyzed for O2 access routes enhanced when the complex forms. Evaluation of the structures of oxidized, diferric Mt sMMOH (sMMOHox, PDB:6VK6, 1.52 A), chemically reduced diferrous Mt sMMOH (sMMOHred, PDB:6VK7, 2.12 A), Mt sMMOHox:MMOB (form 1, PDB ID 6VK5, 1.86 A, and form 2, PDB ID 6VK8, 2.03 A), and Mt sMMOHred:MMOB (PDB ID 6VK4, 2.35 A). The two alphabetagamma protomers of sMMOH protein in the crystal are related by 2fold crystallographic symmetry
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7
sitting drop vapour diffusion method with 100 mM cacodylate, pH 6.5, 20% (v/v) PEG 3000, 250 mM magnesium formate or MnCl2
-
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
A0A2D2D5X0; A0A2D2D0T8; Q53563; A0A2D2D0X7