Visualizing molecular juggling within a B 12-dependent methyltransferase complex

Yan Kung, Nozomi Ando, Tzanko I. Doukov, Leah C. Blasiak, Güneş Bender, Javier Seravalli, Stephen W. Ragsdale, Catherine L. Drennan

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Derivatives of vitamin B 12 are used in methyl group transfer in biological processes as diverse as methionine synthesis in humans and CO 2 fixation in acetogenic bacteria. This seemingly straightforward reaction requires large, multimodular enzyme complexes that adopt multiple conformations to alternately activate, protect and perform catalysis on the reactive B 12 cofactor. Crystal structures determined thus far have provided structural information for only fragments of these complexes, inspiring speculation about the overall protein assembly and conformational movements inherent to activity. Here we present X-ray crystal structures of a complete 220 kDa complex that contains all enzymes responsible for B 12-dependent methyl transfer, namely the corrinoid iron-sulphur protein and its methyltransferase from the model acetogen Moorella thermoacetica. These structures provide the first three-dimensional depiction of all protein modules required for the activation, protection and catalytic steps of B 12-dependent methyl transfer. In addition, the structures capture B 12 at multiple locations between its 'resting' and catalytic positions, allowing visualization of the dramatic protein rearrangements that enable methyl transfer and identification of the trajectory for B 12 movement within the large enzyme scaffold. The structures are also presented alongside in crystallo spectroscopic data, which confirm enzymatic activity within crystals and demonstrate the largest known conformational movements of proteins in a crystalline state. Taken together, this work provides a model for the molecular juggling that accompanies turnover and helps explain why such an elaborate protein framework is required for such a simple, yet biologically essential reaction.

Original languageEnglish (US)
Pages (from-to)265-269
Number of pages5
JournalNature
Volume484
Issue number7393
DOIs
StatePublished - Apr 12 2012

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Methyltransferases
Proteins
Moorella
Enzymes
Corrinoids
Protein Methyltransferases
Iron-Sulfur Proteins
Biological Phenomena
Molecular Models
Carbon Monoxide
Vitamin B 12
Catalysis
Methionine
X-Rays
Bacteria

ASJC Scopus subject areas

  • General

Cite this

Kung, Y., Ando, N., Doukov, T. I., Blasiak, L. C., Bender, G., Seravalli, J., ... Drennan, C. L. (2012). Visualizing molecular juggling within a B 12-dependent methyltransferase complex. Nature, 484(7393), 265-269. https://doi.org/10.1038/nature10916

Visualizing molecular juggling within a B 12-dependent methyltransferase complex. / Kung, Yan; Ando, Nozomi; Doukov, Tzanko I.; Blasiak, Leah C.; Bender, Güneş; Seravalli, Javier; Ragsdale, Stephen W.; Drennan, Catherine L.

In: Nature, Vol. 484, No. 7393, 12.04.2012, p. 265-269.

Research output: Contribution to journalArticle

Kung, Y, Ando, N, Doukov, TI, Blasiak, LC, Bender, G, Seravalli, J, Ragsdale, SW & Drennan, CL 2012, 'Visualizing molecular juggling within a B 12-dependent methyltransferase complex', Nature, vol. 484, no. 7393, pp. 265-269. https://doi.org/10.1038/nature10916
Kung Y, Ando N, Doukov TI, Blasiak LC, Bender G, Seravalli J et al. Visualizing molecular juggling within a B 12-dependent methyltransferase complex. Nature. 2012 Apr 12;484(7393):265-269. https://doi.org/10.1038/nature10916
Kung, Yan ; Ando, Nozomi ; Doukov, Tzanko I. ; Blasiak, Leah C. ; Bender, Güneş ; Seravalli, Javier ; Ragsdale, Stephen W. ; Drennan, Catherine L. / Visualizing molecular juggling within a B 12-dependent methyltransferase complex. In: Nature. 2012 ; Vol. 484, No. 7393. pp. 265-269.
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