Steric Course and Mechanism of Coenzyme B12-Dependent Rearrangements

  • János Rétey
Part of the Industry-University Cooperative Chemistry Program Symposia book series (IUCC)


Vitamin B12 is not only one of the most fascinating naturally occurring molecules, its coenzyme form is also a unique co-catalyst. It is unique both in its structure and in its function. By X-ray diffraction studies of Lehnert and Hodgkin a covalent bond has been identified between the central cobalt atom and its axial ligand, 5′-deoxyadenosine. The stability of this metalorganic bond in aqueous solution is surprising and so is its role in a number of enzymic rearrangements. The first of these, the glutamate mutase reaction was discovered 1958 by H.A. Barker et al.1 (Eqn.1). It was and still is a mechanistically intriguing process without chemical precedence. One of the puzzles for chemists is the specific attack and subsequent substitution at a non-activated methyl group, even though hydrogen atoms in more activated positions are available in the molecule. A similar rearrangement, discovered at about the same time both in bacteria and in mammals2,3, turned out to be also dependent on coenzyme B12 (Eqn.2). In the last few years we focused our attention on this rearrangement catalyzed by the enzyme methylmalonyl-CoA mutase and a considerable portion of my lecture will deal with it. Other coenzyme B12-dependent reactions in which a rearrangement is not immediately obvious are catalyzed by dioldehydratases (Eqn.3) and ammonia lyases (Eqn.4).


Electron Spin Resonance Electron Spin Reso Minimal Mechanism Coenzyme Form Unlabelled Molecule 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    H.A. Barker, H. Weissbach and R.D. Smyth, A coenzyme containing pseudo-vitamin B12, Proc.Natl. Acad. Sci. USA 44:1093 (1958).PubMedCrossRefGoogle Scholar
  2. 2.
    M. Flavin and S. Ochoa, Propionyl coenzyme A carboxylation system, J.Biol.Chem. 229:965 (1957).PubMedGoogle Scholar
  3. 3.
    H. Eggerer, E.R. Stadtman, P. Overath and F. Lynen, Zum Mechanismus der durch Cobalamin-Coenzym katalysierten Umlagerung von Methylmalonyl-CoA in Succinyl-CoA, Biochem. Z. 333:1 (1960).PubMedGoogle Scholar
  4. 4.
    J. Rétey, A. Umani-Ronchi, J. Seibl and D. Arigoni, Zum Mechanismus der Propandioldehydrase-Reaktion, Experientia 22: 502 (1966).PubMedCrossRefGoogle Scholar
  5. 5.
    R.W. Kellermeyer and H.G. Wood, Methylmalonyl-Isomerase, A study of the mechanism of isomerization, Biochemistry 1:112 (1962).CrossRefGoogle Scholar
  6. 6.
    E.F. Phares, M.V. Long and S.F. Carsen, An intramolecular rearrangement in the methylmalonyl isomerase reaction as demonstrated by positive and negative ion mass analysis of succinic acid, Biochem.Biophys. Res. Commun. 8:142 (1962).PubMedCrossRefGoogle Scholar
  7. 7.
    P.A. Frey and R.H. Abeles, The Role of the B12 Coenzyme in the Conversion of 1, 2-Propanediol to Propionaldehyde, J.Biol.Chem. 241:2732 (1966).PubMedGoogle Scholar
  8. 8.
    J. Rétey and D. Arigoni, Coenzym B12 als gemeinsamer Wasserstoffüberträger der Dioldehydrase und der Methylmalonyl-CoA-Mutase-Reaktion, Experientia 22:783 (1966).PubMedCrossRefGoogle Scholar
  9. 9.
    G.J. Cardinale and R.H. Abeles, Mechanistic similarities in the reactions catalyzed by dioldehydrase and methylmalonyl-CoA mutase, Biochim.Biophys.Acta 132:517 (1967).PubMedCrossRefGoogle Scholar
  10. 10.
    D.A. Weisblat and B.M. Babior, Mechanism of action of ethanolamine ammonia-lyase, a B12-dependent enzyme. VIII. Further studies with compounds labeled with isotopes of hydrogen. Identification and some properties of the rate-limiting step, J.Biol.Chem. 246 (19):6064 (1971).PubMedGoogle Scholar
  11. 11.
    P. Diziol, H. Haas, J. Rétey, S.W. Graves and B. Babior, The Substrate-Dependent Steric Course of the Ethanolamine Ammonia-Lyase Reacion, Eur. J. Biochem. 106:211 (1980).PubMedCrossRefGoogle Scholar
  12. 12.
    J. Rétey, F. Kunz, T.C. Stadtman and D. Arigoni, Zum Mechanismus der β-Lysin-Mutase-Reaktion, Experientia 25:801 (1969).PubMedCrossRefGoogle Scholar
  13. 13.
    J. Rétey, F. Kunz, D. Arigoni and T.C. Stadtman, Zur Kenntnis der 3-Lysin-Mutase-Reaktion: Mechanismus und Sterischer Verlauf, Helv.Chim.Acta 61:2989 (1978).CrossRefGoogle Scholar
  14. 14.
    B.M. Babior, T.H. Moss and D.C. Gould, Mechanism of action of ethanolamine ammonia lyase, a B12-dependent enzyme. X. Study of the reaction by electron spin resonance spectrometry, J.Biol.Chem. 247:4389 (1972).PubMedGoogle Scholar
  15. 15.
    M.R. Hollaway, H.A. White,. N. Joblin, A.W. Johnson, M.F. Lappert and O.C. Wallis, Coenzyme-B12-dependent reactions. Part V. A spectrophotometric rapid kinetic study of reactions catalyzed by coenzyme-B12-dependent ethanoloamine ammonia-lyase, Eur. J. Biochem. 82(1):143 (1978).PubMedCrossRefGoogle Scholar
  16. 16.
    B.M. Babior, T.H. Moss, W.H. Orne-Johnson and H. Beinert, Mechanism of action of ethanolamine ammonia-lyase, a B12-dependent enzyme. 13. Participation of paramagnetic species in the catalytic deamination of 2-aminopropanol, J.Biol.Chem. 249(14):4537 (1974).PubMedGoogle Scholar
  17. 17.
    O.C. Wallis, R.C. Bray, S. Gutteridge and M.R. Hollaway, The Extents of Formation of Cobalt(II)-Radical Intermediates in the Reaction with Different Substrates Catalysed by the Adenosylcobalamin-Dependent Enzyme Ethanolamine Ammonia-Lyase, Eur. J. Biochem. 125:299 (1982).PubMedCrossRefGoogle Scholar
  18. 18.
    S.A. Cockle, H.A.O. Hill, R.J.P. Williams, S.P. Davies and M.A. Foster, The Detection of Intermediates During the Conversion of Propane-1, 2-diol to Propionaldehyde by Glyceroldehydrase a Coenzyme B12-Dependent Reaction, J. Am. Chem.Soc. 94:275 (1972).PubMedCrossRefGoogle Scholar
  19. 19.
    K.L. Schepler, W.R. Dunham, R.H. Sands, J.A. Fee and R.H. Abeles, Physical explanation of the EPR spectrum observed during catalysis by enzymes utilizing coenzyme B12, Biochim. Biophys. Acta 397(2):510 (1975).PubMedCrossRefGoogle Scholar
  20. 20.
    G.R. Buettner and R.E. Coffman, EPR Determination of the Co(II)-Free Radical Magnetic Geometry of the “Doublet” Species Arising in a Coenzyme B12 Enzyme Reaction, Biochim. Biophys. Acta 480:495 (1977).PubMedCrossRefGoogle Scholar
  21. 21.
    J.R. Boas, P.R. Hicks, J.R. Pilbrow and T.D. Smith, Interpretation of Electron Spin Resonance Spectra Due to Some B12-dependent Enzyme Reactions, J. Chem.Soc. Faraday II 74:417 (1978).CrossRefGoogle Scholar
  22. 22.
    J. Rétey and J.A. Robinson, Stereospecificity in Organic Chemistry and Enzymology, Verlag Chemie, Weinheim/Deer-field Beach/Basel (1982).Google Scholar
  23. 23.
    J. Rétey, Vitamin B12: Stereochemical aspects of its biological functions and of its biosynthesis, in: “Stereochemistry”, C. Tamm, ed. New Comprehensive Biochemistry, Vol. 3, Elsevier Biomedical Press (1982).Google Scholar
  24. 24.
    J.W. Cornforth, J.W. Redmond, H. Eggerer, W. Buckel and C. Gutschow, Asymmetric methyl groups, Nature 221:1212 (1969).PubMedCrossRefGoogle Scholar
  25. 25.
    J. Lüthy, J. Rétey and D. Arigoni, Preparation and Detection of Chiral Methyl Groups, Nature 221:1213 (1969).PubMedCrossRefGoogle Scholar
  26. 26.
    J. Rétey, A. Umani-Ronchi and D. Arigoni, Zur Stereochemie der Propandioldehydrase-Reaktion, Experientia 22:72 (1966).PubMedCrossRefGoogle Scholar
  27. 27.
    M. Sprecher, M.S. Clark and D.B. Sprinson, The Absolute Configuration of Methylmalonyl-Coenzyme A and Stereochemistry of the Methylmalonyl-Coenzyme A Mutase Reaction, J. Biol. Chem. 241:872 (1966).PubMedGoogle Scholar
  28. 28.
    J. Rétey, cited by D. Arigoni and E.L. Eliel, Chirality Due to the Presence of Hydrogen Isotopes at Noncyclic Positions, in: “Top. Stereochemistry”, vol. 4, E.L. Eliel and N.L. Allinger, eds. J. Wiley and Sons, New York (1969).Google Scholar
  29. 29.
    J. Rétey, E.H. Smith and B. Zagalak, Investigation of the Mechanism of the Methylmalonyl-CoA Mutase Reaction with the Substrate Analogue: Ethylmalonyl-CoA, Eur. J. Biochemistry 83:437 (1978).CrossRefGoogle Scholar
  30. 30.
    K. Wölfle, M. Michenfelder, A. König, W.E. Hull and J. Rétey, On the mechanism of action of methylmalonyl-CoA mutase. Change of the steric course on isotope substitution, Eur. J. Biochem. 156:545 (1986).PubMedCrossRefGoogle Scholar
  31. 31.
    M. Michenfelder, W.E. Hull and J. Rétey, Quantitative measurement of the error in the cryptic stereospecificity of methylmalonyl-CoA mutase, Eur. J. Biochem. 168:659 (1987).PubMedCrossRefGoogle Scholar
  32. 32.
    W.E. Hull, M. Michenfelder and J. Rétey, The error in the cryptic stereospecificity of methylmalonyl-CoA mutase. The use of carba-(dethia)-coenzyme A substrate analogues gives new insight into the enzyme mechanism, Eur. J. Biochem. 173:191 (1988).PubMedCrossRefGoogle Scholar
  33. 33.
    K. Aeberhard, R. Keese, E. Stamm, V.R. Vögel, W. Lau and J. Kochi, Structure and Chemistry of Malonylmethyl-and Succinyl-Radicals. The Search for Homolytic 1,2-Rearrangements, Helv.Chim. Acta 66:2740 (1983).CrossRefGoogle Scholar
  34. 34.
    S. Wollowitz and J. Halpern, 1,2-Migrations in Free Radicals Related to Coenzyme B12-Dependent Rearrangements, J. Am. Chem. Soc. 110:3112 (1988).CrossRefGoogle Scholar
  35. 35.
    W. Best and D.A. Widdowson, Reactions Related to Coenzyme B12 Dependent Rearrangements: Metal Mediated Free Radical Acyl Migrations in Methyl and Cyclopropyl Substituted Models, Tetrahedron 45:5943 (1989).CrossRefGoogle Scholar
  36. 36.
    P. Dowd, The Vitamin B12 Promoted Model Rearrangement of Methylmalonate to Succinate is Not a Free Radical Reaction, Contribution to these Proceedings (1990).Google Scholar
  37. 37.
    F.D. Ledley, M. Lumetta, P.N. Nguyen, J.F. Kolhouse and R.H. Allen, Molecular cloning of L-methylmalonyl-CoA Mutase: Gene transfer and analysis of mut cell lines, Proc.Natl.Acad.Sci. USA, 85:3518 (1988).PubMedCrossRefGoogle Scholar
  38. 38.
    R. Jansen, F. Kalousek, W.A. Fenton, L.E. Rosenberg and F.D. Ledley, Cloning of Full-Length Methylmalonyl-CoA Mutase from a cDNA Library Using the Polymerase Chain Reaction, Genomics 4:198 (1989).PubMedCrossRefGoogle Scholar
  39. 39.
    R.W. Kellermeyer and H.G. Wood, 2-Methylmalonyl-CoA Mutase from Propionibacterium shermanii (Methylmalonyl-CoA Isomerase), Methods Enzymol. 13:207 (1969).CrossRefGoogle Scholar
  40. 40.
    B. Zagalak, J. Rétey and H. Sund, Studies on Methylmalo-nyl-CoA Mutase from Propionibacterium shermanii, Eur. J. Biochem. 44:529 (1974).PubMedCrossRefGoogle Scholar
  41. 41.
    F. Francalanci, N.K. Davis, J.K. Fuller, D.M. Murfitt and P.F. Leadlay, The subunit structure of methylmalonyl-CoA mutase from Propionibacterium shermanii, Biochem. J. 236:489 (1986).PubMedGoogle Scholar
  42. 42.
    E.N. Marsh, N. McKie, N.K. Davis and P.F. Leadlay, Cloning and structural characterization of the genes coding for adenosylcobalamin-dependent methylmalonyl-CoA mutase from Propionibacterium shermanii, Biochem. J. 260:345 (1989).PubMedGoogle Scholar
  43. 43.
    I. Burkhart, J. Kleiber, R. Müller, G. Reck and J. Rétey, Investigation of Methylmalonyl-CoA Mutase from Several Sources by Gene Technological Methods, Biol. Chem. Hoppe-Seyler 368:1028 (1987).Google Scholar

Copyright information

© Springer Science+Business Media New York 1990

Authors and Affiliations

  • János Rétey
    • 1
  1. 1.Chair of BiochemistryUniversity of KarlsruheGermany

Personalised recommendations