Spectroscopic and Kinetic Studies of the Reaction of Bromopropanesulfonate with Methyl-coenzyme M Reductase

Kunz, Ryan C.; Horng, Yih-Chern; Ragsdale, Stephen W.

doi:10.1074/jbc.m606715200

Cited by 35 publications

(104 citation statements)

References 51 publications

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“…M. marburgensis was cultured on H 2 /CO 2 (80/ 20%) at 65°C in a 14-liter fermentor (New Brunswick Scientific Co., Inc., New Brunswick, NJ) to an optical density of 5-6 at 578 nm. Culture media were prepared as previously described (39) with a slight modification of the sulfur and reducing source, by adding 50 mM sodium sulfide (instead of H 2 S gas) at a flow rate of 1 ml/min during the entire growth period. Before harvest, the cells were treated with 100% H 2 for 30 min in the fermenter.…”

Section: Methodsmentioning

confidence: 99%

“…The headspace of the bottle containing the resuspended cells was then purged with CO for 10 min or at timed intervals to generate the active MCR red1 state as previously described (17). The purification of MCR red1 (Ni(I) state) was performed as described earlier (39). All steps were performed in the presence of 10 mM HSCoM and 0.1 mM Ti(III) citrate.…”

Section: Methodsmentioning

confidence: 99%

“…This purification procedure generates about 60 -70% MCR red1 and this is the form of the enzyme that was used in all experiments unless otherwise stated. The concentration of MCR red1 was determined by UV-visible spectroscopy using extinction coefficients of 27.0 and 9.15 mM Ϫ1 cm Ϫ1 at 385 and 420 nm, respectively, using a multiple wavelength calculation as previously described (39). The concentration of MCR silent , which contains the inactive Ni(II) form of F430, was calculated using extinction coeffi-cients of 22.0 and 12.7 mM Ϫ1 cm Ϫ1 at 420 and 385 nm, respectively (39).…”

Section: Methodsmentioning

confidence: 99%

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The Reaction Mechanism of Methyl-Coenzyme M Reductase

Wongnate

Ragsdale

2015

Journal of Biological Chemistry

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Background: Methyl-coenzyme M reductase (MCR) catalyzes the final step in methanogenesis. Results: MCR forms binary complexes with both substrates, but stabilizes only the productive binary and ternary complexes. Conclusion: Substrate-induced conformational changes promote correct binding order and chemistry. Significance: This first rapid kinetics study of MCR with natural substrates describes how an enzyme can enforce a strictly ordered ternary complex reaction mechanism.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The Reaction Mechanism of Methyl-Coenzyme M Reductase

Wongnate

Ragsdale

2015

Journal of Biological Chemistry

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“…At the ''single-crystal'' observer position corresponding to g 3 (Fig. 2c, 1,098 mT) [53], MCR BPS [14,15], and MCR ox1 [54], but vastly different from those of MCR red2r [53], where the four F 430 hydropyrrolic nitrogen hyperfine interactions span a large range, indicating a significant asymmetry in the spin-density distribution (this species has a strong proximal sulfur ligand from HS-CoM). The outcome for MCR red1m is that the spin-density distribution on F 430 is not significantly perturbed by the weak axial ligand.…”

Section: Hydropyrrolic Nitrogens Of F 430mentioning

confidence: 99%

“…in the active site [14][15][16][17], with Br -/I -as the leaving group. Additional evidence is provided by the reaction of free Ni(I)F 430 derivatives with electrophilic methyl donors such as methyl-dialkylsulfonium ions and methyl halides [16,17].…”

Section: -Ni(iii)f 430mentioning

confidence: 99%

Coordination and binding geometry of methyl-coenzyme M in the red1m state of methyl-coenzyme M reductase

et al. 2008

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Methane formation in methanogenic Archaea is catalyzed by methyl-coenzyme M reductase (MCR) and takes place via the reduction of methyl-coenzyme M (CH 3 -S-CoM) with coenzyme B (HS-CoB) to methane and the heterodisulfide CoM-S-S-CoB. MCR harbors the nickel porphyrinoid coenzyme F 430 as a prosthetic group, which has to be in the Ni(I) oxidation state for the enzyme to be active. To date no intermediates in the catalytic cycle of MCR red1 (red for reduced Ni) have been identified. Here, we report a detailed characterization of MCR red1m (''m'' for methyl-coenzyme M), which is the complex of MCR red1a (''a'' for absence of substrate) with CH 3 -S-CoM. Using continuous-wave and pulse electron paramagnetic resonance spectroscopy in combination with selective isotope labeling ( 13 C and 2 H) of CH 3 -S-CoM, it is shown that CH 3 -S-CoM binds in the active site of MCR such that its thioether sulfur is weakly coordinated to the Ni(I) of F 430 . The complex is stable until the addition of the second substrate, HS-CoB. Results from EPR spectroscopy, along with quantum mechanical calculations, are used to characterize the electronic and geometric structure of this complex, which can be regarded as the first intermediate in the catalytic mechanism.

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Biochemistry of Methyl-Coenzyme M Reductase: The Nickel Metalloenzyme that Catalyzes the Final Step in Synthesis and the First Step in Anaerobic Oxidation of the Greenhouse Gas Methane

Ragsdale

2014

The Metal-Driven Biogeochemistry of Gaseous Compounds in the Environment

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Methane, the major component of natural gas, has been in use in human civilization since ancient times as a source of fuel and light. Methanogens are responsible for synthesis of most of the methane found on Earth. The enzyme responsible for catalyzing the chemical step of methanogenesis is methyl-coenzyme M reductase (MCR), a nickel enzyme that contains a tetrapyrrole cofactor called coenzyme F430, which can traverse the Ni(I), (II), and (III) oxidation states. MCR and methanogens are also involved in anaerobic methane oxidation. This review describes structural, kinetic, and computational studies aimed at elucidating the mechanism of MCR. Such studies are expected to impact the many ramifications of methane in our society and environment, including energy production and greenhouse gas warming.

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Spectroscopic and Kinetic Studies of the Reaction of Bromopropanesulfonate with Methyl-coenzyme M Reductase

Cited by 35 publications

References 51 publications

The Reaction Mechanism of Methyl-Coenzyme M Reductase

The Reaction Mechanism of Methyl-Coenzyme M Reductase

Coordination and binding geometry of methyl-coenzyme M in the red1m state of methyl-coenzyme M reductase

Biochemistry of Methyl-Coenzyme M Reductase: The Nickel Metalloenzyme that Catalyzes the Final Step in Synthesis and the First Step in Anaerobic Oxidation of the Greenhouse Gas Methane

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