CofE Catalyzes the Addition of Two Glutamates to F420-0 in F420 Coenzyme Biosynthesis in Methanococcus jannaschii

Li, Hong; Graupner, Marion; Xu, Huimin; White, Robert H.

doi:10.1021/bi034779b

Cited by 55 publications

(68 citation statements)

References 35 publications

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“…Subsequently, LPPG (L-lactyl-2-diphospho-5 0 -guanosine) is proposed to be synthesized from 2-phospho-L-lactate by CofC (Grochowski et al, 2008) and transferred to F o by CofD (also known as FbiA) (Choi et al, 2001;Graupner and White, 2001;Graupner et al, 2002). The resulting LPPG sidechain is finally elongated with glutamate residues by the F 420 :γ-L-glutamyl ligase CofE (also known as FbiB) (Choi et al, 2001;Li et al, 2003;Nocek et al, 2007) that is fused with an FMNdependent oxidoreductase in Actinobacteria (Bashiri et al, 2016). For reasons still not understood, the number of glutamate residues added varies between organisms, ranging from two to three in most methanogens (Gorris and van der Drift, 1994), four to five in Methanosarcina (Gorris and van der Drift, 1994) and five to seven in Mycobacterium .…”

Section: Introductionmentioning

confidence: 99%

The methanogenic redox cofactor F420 is widely synthesized by aerobic soil bacteria

et al. 2016

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F 420 is a low-potential redox cofactor that mediates the transformations of a wide range of complex organic compounds. Considered one of the rarest cofactors in biology, F 420 is best known for its role in methanogenesis and has only been chemically identified in two phyla to date, the Euryarchaeota and Actinobacteria. In this work, we show that this cofactor is more widely distributed than previously reported. We detected the genes encoding all five known F 420 biosynthesis enzymes (cofC, cofD, cofE, cofG and cofH) in at least 653 bacterial and 173 archaeal species, including members of the dominant soil phyla Proteobacteria, Chloroflexi and Firmicutes. Metagenome datamining validated that these genes were disproportionately abundant in aerated soils compared with other ecosystems. We confirmed through high-performance liquid chromatography analysis that aerobically grown stationary-phase cultures of three bacterial species, Paracoccus denitrificans, Oligotropha carboxidovorans and Thermomicrobium roseum, synthesized F 420 , with oligoglutamate sidechains of different lengths. To understand the evolution of F 420 biosynthesis, we also analyzed the distribution, phylogeny and genetic organization of the cof genes. Our data suggest that although the F o precursor to F 420 originated in methanogens, F 420 itself was first synthesized in an ancestral actinobacterium. F 420 biosynthesis genes were then disseminated horizontally to archaea and other bacteria. Together, our findings suggest that the cofactor is more significant in aerobic bacterial metabolism and soil ecosystem composition than previously thought. The cofactor may confer several competitive advantages for aerobic soil bacteria by mediating their central metabolic processes and broadening the range of organic compounds they can synthesize, detoxify and mineralize.

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Section: Introductionmentioning

confidence: 99%

The methanogenic redox cofactor F420 is widely synthesized by aerobic soil bacteria

et al. 2016

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“…A range of different conditions was optimized for the ␥-glutamyl ligase activity of the full-length FbiB protein, including pH (6.0 -9.0), monovalent (Na ϩ and K ϩ ) and divalent cation (Mg 2ϩ and Mn 2ϩ ) composition, nucleotides (GTP, dGTP, ATP, and dATP), and also various time points and temperatures. FbiB showed the highest activity at pH 8.5, the same pH dependence displayed by the CofE protein (17). A combination of Na ϩ and Mn 2ϩ produced the highest activity in FbiB, in contrast to the previously reported dependence on K ϩ and (17).…”

Section: Resultsmentioning

confidence: 53%

“…FbiB showed the highest activity at pH 8.5, the same pH dependence displayed by the CofE protein (17). A combination of Na ϩ and Mn 2ϩ produced the highest activity in FbiB, in contrast to the previously reported dependence on K ϩ and (17). The enzyme was only active in the presence of GTP, with no observed activity for dGTP, ATP, and dATP nucleotides.…”

Section: Resultsmentioning

confidence: 58%

“…The ␥-glutamyl ligase activity of FbiB constructs was investigated by monitoring the addition of L-glutamate residues to F 420 -0 using HPLC, as described previously for the CofE protein (16). The F 420 -0 substrate was prepared by enzymatic hydrolysis of the poly-␥-glutamate tail of F 420 using carboxypeptidase G (17), and the resulting F 420 -0 was used as a substrate for FbiB activity experiments and also for crystallographic binding studies.…”

Section: Resultsmentioning

confidence: 99%

“…The optimized reaction mixture included 50 mM HEPES, pH 8.5, 100 mM NaCl, 5 mM MnCl 2 , 10 mM L-glutamate, 5 mM GTP, and 2 M F 420 -0. The F 420 -0 substrate was prepared by enzymatic hydrolysis of the poly-␥-glutamate tail of F 420 using carboxypeptidase G (in 50 mM Tris-HCl, pH 7.5, 0.1 M NaCl, 0.2 mM ZnSO 4 ) (17). The possible effect of FMN on the polyglutamylation reaction was also tested by including a range of FMN concentrations (0 -1 mM) in duplicate activity assays.…”

Section: Pcr Amplification and Cloning-mentioning

confidence: 99%

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Elongation of the Poly-γ-glutamate Tail of F420 Requires Both Domains of the F420:γ-Glutamyl Ligase (FbiB) of Mycobacterium tuberculosis

Bashiri

Rehan

Sreebhavan

et al. 2016

Journal of Biological Chemistry

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Cofactor F 420 is an electron carrier with a major role in the oxidoreductive reactions of Mycobacterium tuberculosis, the causative agent of tuberculosis. A ␥-glutamyl ligase catalyzes the final steps of the F 420 biosynthesis pathway by successive additions of L-glutamate residues to F 420 -0, producing a poly-␥-glutamate tail. The enzyme responsible for this reaction in archaea (CofE) comprises a single domain and produces F 420 -2 as the major species. The homologous M. tuberculosis enzyme, FbiB, is a two-domain protein and produces F 420 with predominantly 5-7 L-glutamate residues in the poly-␥-glutamate tail. The N-terminal domain of FbiB is homologous to CofE with an annotated ␥-glutamyl ligase activity, whereas the C-terminal domain has sequence similarity to an FMN-dependent family of nitroreductase enzymes. Here we demonstrate that full-length FbiB adds multiple L-glutamate residues to F 420 -0 in vitro to produce F 420 -5 after 24 h; communication between the two domains is critical for full ␥-glutamyl ligase activity. We also present crystal structures of the C-terminal domain of FbiB in apo-, F 420 -0-, and FMN-bound states, displaying distinct sites for F 420 -0 and FMN ligands that partially overlap. Finally, we discuss the features of a full-length structural model produced by small angle x-ray scattering and its implications for the role of N-and C-terminal domains in catalysis.The cofactor F 420 is a flavin derivative that is sporadically distributed among microorganisms, mainly archaea and actinobacteria (including mycobacteria). F 420 has been emerging as a new player in the biology of mycobacteria (1), with increasing numbers of F 420 -utilizing proteins characterized from different mycobacterial species (2-8). This cofactor has been suggested to protect Mycobacterium tuberculosis, the causative agent of tuberculosis, against oxidative and nitrosative stress during pathogenesis (9 -11). At the biochemical level, cofactor F 420 functions as a hydride transfer agent in oxidoreductive reactions with a lower redox potential than that of NAD(P) ϩ (12). The biosynthesis pathway of cofactor F 420 has been investigated in both archaeal and mycobacterial species. In the current view of the proposed pathway, the first intermediate with the complete chromophore (7, ) is produced by FO synthase (FbiC in mycobacteria (13) and CofGH in archaea (14)). A transferase enzyme (FbiA in mycobacteria (15) and CofD in archaea (16)) subsequently catalyzes the addition of a 2-phospho-L-lactate moiety to FO to produce F 420 -0 (F 420 with no poly-␥-glutamate tail). The final step of the pathway is performed by a ␥-glutamyl ligase (FbiB in mycobacteria (15) and CofE in archaea (17)) that catalyzes successive additions of L-glutamate residues to F 420 -0 (Fig. 1A).The length of the poly-␥-glutamate tail varies between archaeal and mycobacterial species; in archaea, two L-glutamate residues are seen (18), whereas in mycobacteria, up to nine residues are present (3, 19). There exists an intriguing difference between the ...

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coenzyme F420-0:l-glutamate ligase 6.3.2.31

Schomburg¹,

Schomburg²

2013

Class 3.4–6 Hydrolases, Lyases, Isomerases, Ligases

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CofE Catalyzes the Addition of Two Glutamates to F₄₂₀-0 in F₄₂₀ Coenzyme Biosynthesis in Methanococcus jannaschii

Cited by 55 publications

References 35 publications

The methanogenic redox cofactor F420 is widely synthesized by aerobic soil bacteria

The methanogenic redox cofactor F420 is widely synthesized by aerobic soil bacteria

Elongation of the Poly-γ-glutamate Tail of F420 Requires Both Domains of the F420:γ-Glutamyl Ligase (FbiB) of Mycobacterium tuberculosis

coenzyme F420-0:l-glutamate ligase 6.3.2.31

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