Elucidation of the coenzyme binding mode of further B12-dependent enzymes using a base-off analogue of coenzyme B12

L. Poppe, Harald Bothe, Gerd Bröker, Wolfgang Buckel, Erhard Stupperich, János Rétey

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

(Coβ-5'-Deoxyadenosin-5'-yl)-(p-cresyl)cobamide (Ado-PCC), a base-off analogue of coenzyme B12 (Ado-Cbl), was used to elucidate the coenzyme B12 binding mode of glutamate mutase, 2-methyleneglutarate mutase and ethanolamine ammonia-lyase (EAL). Ado-PCC functions as excellent coenzyme for the carbon skeleton rearrangements with apparent K(m) values of 200 and 10 nM for glutamate and 2-methyleneglutarate mutases, respectively. The corresponding values for Ado-Cbl are 60 and 54 nM, respectively. In contrast, Ado-PCC showed no coenzyme activity with EAL but was a competitive inhibitor with respect to Ado-Cbl. The K(i) value for Ado-PCC was 26 nM, the apparent K(m) value for Ado-Cbl was 30 nM. These results are in agreement with the notion that in glutamate and 2-methyleneglutarate mutases, a conserved histidine residue of the protein is coordinated to the cobalt atom of coenzyme B12, whereas in EAL the dimethylbenzimidazole residue of the coenzyme itself serves as ligand. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)345-350
Number of pages6
JournalJournal of Molecular Catalysis B: Enzymatic
Volume10
Issue number1-3
DOIs
Publication statusPublished - Oct 4 2000

Fingerprint

Ethanolamine Ammonia-Lyase
Coenzymes
methylaspartate mutase
Enzymes
Ethanolamines
Glutamic Acid
Cobamides
Ammonia
Cobalt
Histidine
Skeleton
Carbon
Ligands
Atoms
cobamamide
2-methyleneglutarate mutase
Proteins

Keywords

  • (Coβ-5'-deoxyadenosin-5'-yl)-(p-cresyl)cobamide
  • 2-Methyleneglutarate mutase (Clostridium barkeri)
  • Base-off analogue of coenzyme B
  • Ethanolamine ammonia lyase (Salmonella typhimurium)
  • Glutamate mutase (Clostridium cochlearium)

ASJC Scopus subject areas

  • Biochemistry
  • Catalysis
  • Process Chemistry and Technology

Cite this

Elucidation of the coenzyme binding mode of further B12-dependent enzymes using a base-off analogue of coenzyme B12 . / Poppe, L.; Bothe, Harald; Bröker, Gerd; Buckel, Wolfgang; Stupperich, Erhard; Rétey, János.

In: Journal of Molecular Catalysis B: Enzymatic, Vol. 10, No. 1-3, 04.10.2000, p. 345-350.

Research output: Contribution to journalArticle

Poppe, L. ; Bothe, Harald ; Bröker, Gerd ; Buckel, Wolfgang ; Stupperich, Erhard ; Rétey, János. / Elucidation of the coenzyme binding mode of further B12-dependent enzymes using a base-off analogue of coenzyme B12 In: Journal of Molecular Catalysis B: Enzymatic. 2000 ; Vol. 10, No. 1-3. pp. 345-350.
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