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Domain alternation switches B12-dependent methionine synthase to the activation conformation

Nature Structural Biology volume 9, pages 53–56 (2002)Cite this article

Abstract

B12-dependent methionine synthase (MetH) from Escherichia coli is a large modular protein that uses bound cobalamin as an intermediate methyl carrier. Major domain rearrangements have been postulated to explain how cobalamin reacts with three different substrates: homocysteine, methyltetrahydrofolate and S-adenosylmethionine (AdoMet). Here we describe the 3.0 Å structure of a 65 kDa C-terminal fragment of MetH that spans the cobalamin- and AdoMet-binding domains, arranged in a conformation suitable for the methyl transfer from AdoMet to cobalamin that occurs during activation. In the conversion to the activation conformation, a helical domain that capped the cofactor moves 26 Å and rotates by 63°, allowing formation of a new interface between cobalamin and the AdoMet-binding (activation) domain. Interactions with the MetH activation domain drive the cobalamin away from its binding domain in a way that requires dissociation of the axial cobalt ligand and, thereby, provide a mechanism for control of the distribution of enzyme conformations.

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Figure 1: Methionine synthesis and reactivation of methionine synthase.
Figure 2: A stereo ribbon drawing of the C-terminal (651–1227) fragment of methionine synthase.
Figure 3: Displacement of the cobalamin and changes in its interactions with the protein.
Figure 4: Stereo views of electron density in the vicinity of the cobalamin and the AdoMet binding site.

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Acknowledgements

Supported by NIH grants to M.L.L. and to R.G.M., and NRSA postdoctoral fellowships to V.B. and B.W.L. are acknowledged.

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  1. Biophysics Research Division and Department of Biological Chemistry, University of Michigan, Ann Arbor, 48109-1055, Michigan, USA

    Vahe Bandarian, Katherine A. Pattridge, Brett W. Lennon, Donald P. Huddler, Rowena G. Matthews & Martha L. Ludwig

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  1. Vahe Bandarian
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  2. Katherine A. Pattridge
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  3. Brett W. Lennon
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  4. Donald P. Huddler
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  5. Rowena G. Matthews
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Correspondence to Martha L. Ludwig.

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Bandarian, V., Pattridge, K., Lennon, B. et al. Domain alternation switches B12-dependent methionine synthase to the activation conformation. Nat Struct Mol Biol 9, 53–56 (2002). https://doi.org/10.1038/nsb738

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