Cloning and sequencing of glutamate mutase component S from <i>Clostridium tetanomorphum</i> Homologies with other cobalamin‐dependent enzymes

Marsh, E. Neil G.; Holloway, Daniel E.

doi:10.1016/0014-5793(92)81321-c

Cited by 142 publications

(122 citation statements)

References 22 publications

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“…Several amino acid residues are conserved through all these 10 protein sequences including a DXHXXG motif at D 484 in Mgm (Fig. 4) that was previously reported by Marsh and Holloway (1992).…”

Section: Amino Acid Sequence Similaritiessupporting

confidence: 55%

“…Mgm aligned not only with the C-terminal part of all known methylmalonyl-CoA mutase sequences as are that of the human (Jansen et a]., 1989), mouse (Nham, 1990), Propionibacterium shermaizii and P. freudenreichii (Marsh et al, 1989) and Streptomyces cinnamonensis (Birch et al, 1993) enzymes but also with glutamate mutase (component S) of Clostridium tetanomorphum (Marsh and Holloway, 1992) and C. cochlearium (Zelder, unpublished results), with the SBM protein of E. coli (unpublished results) and with the cobalamin-dependent methionine synthase (MetH) from E. coEi (Banerjee et al, 1989 ; Fig. 4).…”

Section: Amino Acid Sequence Similaritiesmentioning

confidence: 91%

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Cloning, sequencing and expression of the gene encoding the coenzyme B₁₂‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Beatrix

Zelder

Linder

et al. 1994

European Journal of Biochemistry

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The coenzyme B,, (adenosy1cobalamin)-dependent 2-methyleneglutarate mutase catalyses the carbon skeleton rearrangement of 2-methyleneglutarate to (R)-3-methylitaconate in the fermentation of nicotinic acid by the strict anaerobic bacterium Clostridium barkeri.(a) The mgm gene encoding 2-methyleneglutarate mutase was cloned and its nucleotide sequence was determined. The deduced amino acid sequence revealed a 66.8-kDa protein of 614 amino acids. It shows significant similarity in its C-terminal part to that of other cobamide-dependent enzymes. Probably, this is the coenzyme-binding region.(b) The mgm gene from C. barkeri was expressed in Escherichia coli as was shown by SDS/ PAGE and Westem-blot analysis with rabbit antiserum directed against the native mutase.(c) Cell-free extracts from E. coli carrying the mgm gene showed 2-methyleneglutarate mutase activity that was strictly dependent on the addition of coenzyme B,2. Experiments are presented which suggest that the expression product is an apoenzyme.

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Section: Amino Acid Sequence Similaritiessupporting

confidence: 55%

Section: Amino Acid Sequence Similaritiesmentioning

confidence: 91%

Cloning, sequencing and expression of the gene encoding the coenzyme B₁₂‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Beatrix

Zelder

Linder

et al. 1994

European Journal of Biochemistry

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“…In contrast, glycerol dehydratase and diol dehydratase seem to belong to the 'base-on' family. No similarity with the characteristic sequence pattern [18] of known 'base-off' enzymes, considering the conserved residues that participate in The inhibition patterns of C,-C, analogues on glycerol dehydratase and diol dehydratase alternate in a similar manner: analogues with odd numbers of inserted methylene units seem to be slightly stronger inhibitors than those with even numbers of methylenes. Such alternation was found in the orientation of adenosyl moiety relative to the corrin part in the calculated conformations of C,-C, compounds, for 'H-NMR chemical shifts of several signals of these analogues, or for their solubilities [8].…”

Section: Resultsmentioning

confidence: 99%

Kinetic Investigations with Inhibitors that Mimic the Postomolysis Intermediate in the Reactions of Coenzyme‐B₁₂‐Dependent Glycerol Dehydratase and Diol Dehydratase

Poppe¹,

Rétey²

1997

European Journal of Biochemistry

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Kinetic investigations were performed on the coenzyme-B ,,-dependent glycerol dehydratase and diol dehydratase reactions using 1,2-propanediol as substrate and [o-(adenosin-5'-O-yl)alkyl]cobalamins as mimics of the posthomolysis intermediate state of the coenzyme. All the coenzyme-B,, analogues with oligomethylene chains (C,-C,) inserted between the central Co atom and the 5' 0 of the adenosine moiety were competitive inhibitors with respect to coenzyme B,2. The apparent inhibition constants (K,) of the shorter-chain inhibitors, especially the C, inhibitor, were smaller for both enzymes than those of the longer-chain (Cn, C,) compounds. These results are in agreement with the expected (0.6-0.9 nm) distance between the Co and 5'-methylene paramagnetic centers in the posthomolysis intermediate state of coenzyme B12 in these reactions. Coenzyme B,? (CoBI2) is an essential cofactor for several enzymes that catalyze rearrangement reactions [ 1-41. This molecule contains a unique covalent Co-C bond, which is stable in aqueous solution but easily undergoes homolytic cleavage in COB ,2-dependent enzymatic reactions. This homolysis is the essential initial step in the catalytic cycles of all COB,,-dependent enzymatic processes. Binding of COB,? to the apoprotein provides energy for the Co-C bond cleavage and protects the highly reactive intermediates from the environment [ S ] . COB ,2-dependent enzymes promote cobalt-carbon bond cleavage by application of a stretching force by interaction with the adenosyl moiety and corrin part 161. The energy required for this stretching is provided by a conformational change of the apoprotein, which is probably triggered by binding the substrate. In the posthomolysis complex, however, the protein should have in a catalytically active, relaxed conformation.Therefore, coenzyme-B , 2 analogues that mimic the geometry of the posthomolysis intermediate, i.e. containing both the corrin part and the adenosyl moiety at a distance that corresponds to the distance between these moieties in the COB,, . apoenzyme complex after the homolytic Co-C bond cleavage, may provide valuable information on the structure of the activated complex.For example, the best fitting analogue is expected to be the strongest inhibitor of a given COB ,?-dependent reaction. A series of such posthomolysis-intermediate analogues (Fig. I) were vaned by insertion of an oligomethylene chain (C,-C,) between the 5' 0 of adenosine and the Co of corrin. has been therefore synthesized [7].Based on the principle of 'negative catalysis' [5] and the space requirement of the substrate, we have postulated a distance of approximately 1 nm between ColI and the 5' C radical in the posthomolysis intermediate of the (R)-methylmalonyl-CoA mutase from Propionibucterium shermanii, and we have tested our hypothesis by studying the inhibitory properties of these analogues [8]. Apparent inhibition constants for the C,-C, analogues showed the predicted trend : the strongest inhibition was found with the C, analogue, in which, assuming a zig-zag...

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“…In contrast, the crystal structure of a diol dehydratase showed the bound coenzyme B 12 in its "classic" base-on constitution (63). Accordingly, questions concerning the effect of the nucleotide base on the binding the B 12 cofactors by their apoproteins (35,53,69) and on the mechanisms of B 12 -dependent enzymatic reactions have received more attention (15,43).…”

mentioning

confidence: 99%

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A

et al. 2000

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The corrinoids from the obligate anaerobe Clostridium cochlearium were extracted as a mixture of Co ␤ -cyano derivatives. From 50 g of frozen cells, approximately 2 mg (1.5 mol) of B 12 derivatives was obtained as a crystalline sample. Analysis of the corrinoid sample of C. cochlearium by a combination of high-pressure liquid chromatography and UV-Vis absorbance spectroscopy revealed the presence of three cyano corrinoids in a ratio of about 3:1:1. The spectroscopic data acquired for the sample indicated the main components to be pseudovitamin B 12 (Co ␤ -cyano-7؆-adeninylcobamide) (60%) and factor A (Co ␤ -cyano-7؆-[2-methyl]adeninylcobamide) (20%). Authentic pseudovitamin B 12 was prepared by guided biosynthesis from cobinamide and adenine. Both pseudovitamin B 12 and its homologue, factor A, were subjected to complete spectroscopic analysis by UV-Vis, circular dichroism, mass spectrometry, and by one-and two-dimensional 1 H, 13 C-, and 15 N nuclear magnetic resonance (NMR) spectroscopy. The third component was indicated by the mass spectra to be an isomer of factor A and is likely (according to NMR) to be 7؆-[N 6 -methyl]-adeninylcobamide, a previously unknown corrinoid. C. cochlearium thus biosynthesizes as its native "complete" B 12 cofactors the 7؆-adeninylcobamides and two homologous corrinoids, in which the nucleotide base is a methylated adenine.

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Cloning and sequencing of glutamate mutase component S from Clostridium tetanomorphum Homologies with other cobalamin‐dependent enzymes

Cited by 142 publications

References 22 publications

Cloning, sequencing and expression of the gene encoding the coenzyme B₁₂‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Cloning, sequencing and expression of the gene encoding the coenzyme B₁₂‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Kinetic Investigations with Inhibitors that Mimic the Postomolysis Intermediate in the Reactions of Coenzyme‐B₁₂‐Dependent Glycerol Dehydratase and Diol Dehydratase

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A

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Cloning and sequencing of glutamate mutase component S from Clostridium tetanomorphum Homologies with other cobalamin‐dependent enzymes

Cited by 142 publications

References 22 publications

Cloning, sequencing and expression of the gene encoding the coenzyme B12‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Cloning, sequencing and expression of the gene encoding the coenzyme B12‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Kinetic Investigations with Inhibitors that Mimic the Postomolysis Intermediate in the Reactions of Coenzyme‐B12‐Dependent Glycerol Dehydratase and Diol Dehydratase

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B12and Factor A

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Cloning, sequencing and expression of the gene encoding the coenzyme B₁₂‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Cloning, sequencing and expression of the gene encoding the coenzyme B₁₂‐dependent 2‐methyleneglutarate mutase from Clostridium barkeri in Escherichia coli

Kinetic Investigations with Inhibitors that Mimic the Postomolysis Intermediate in the Reactions of Coenzyme‐B₁₂‐Dependent Glycerol Dehydratase and Diol Dehydratase

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A