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Crystal structure of 2-oxoisovalerate and dehydrogenase and the architecture of 2-oxo acid dehydrogenase multienzyme complexes

Nature Structural Biology volume 6, pages 785–792 (1999)Cite this article

Abstract

The family of giant multienzyme complexes metabolizing pyruvate, 2-oxoglutarate, branched-chain 2-oxo acids or acetoin contains several of the largest and most sophisticated protein assemblies known, with molecular masses between 4 and 10 million Da. The principal enzyme components, E1, E2 and E3, are present in numerous copies and utilize multiple cofactors to catalyze a directed sequence of reactions via substrate channeling. The crystal structure of a heterotetrameric (α2β2) E1, 2-oxoisovalerate dehydrogenase from Pseudomonas putida, reveals a tightly packed arrangement of the four subunits with the β2-dimer held between the jaws of a 'vise' formed by the α2-dimer. A long hydrophobic channel, suitable to accommodate the E2 lipoyl-lysine arm, leads to the active site, which contains the cofactor thiamin diphosphate (ThDP) and an inhibitor-derived covalent modification of a histidine side chain. The E1 structure, together with previous structural information on E2 and E3, completes the picture of the shared architectural features of these enormous macromolecular assemblies.

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Figure 1: Experimental electron density.
Figure 2: Stereo views of the E1b subunits.
Figure 3: Structural organization of the heterotetrameric P. putida E1 α2β2.
Figure 4: a, Binding of the cofactor thiamin diphosphate at the active site of P. putida E1b.
Figure 5: The lipoyl-lysine arm of the E2 lipoyl domain visiting the active site of P. putida E1b.
Figure 6: Structural composite of the components of a 2-oxo acid dehydrogenase multienzyme complex and the dynamics of its functional cycle.

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Acknowledgements

We thank the members of the Biomolecular Structure Center for assistance and support, in particular S.S. Antonysamy, F. Athappilly, E. Merritt, E. Pohl, M. Redinbo and S. Sarfaty. Access to synchrotron sources SSRL (7.1), CHESS (F1) and ESRF (BM14) is deeply appreciated, and we thank the staff for their assistance. Postdoctoral grant from The Swedish Foundation for International Cooperation in Research and Higher Education (STINT) to A.Æ. is gratefully acknowledged. W.G.J.H. acknowledges a major equipment grant from the Murdock Charitable Trust to the Biomolecular Structure Center. This research was supported by grants from NIH and Presbyterian Health Foundation to J.R.S.

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Authors and Affiliations

  1. Departments of Biological Structure and Biochemistry, Biomolecular Structure Center, University of Washington, P.O. Box 357742, Seattle, 98195-7742 , Washington, USA

    Arnthor Ævarsson, Stewart Turley & Wim G.J. Hol

  2. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, P.O. Box 26901, Oklahoma City, 73190, Oklahoma, USA

    Kimberlie Seger & John R. Sokatch

  3. Howard Hughes Medical Institute, School of Medicine, University of Washington, P.O. Box 357742, Seattle , 98195-7742, Washington, USA

    Stewart Turley & Wim G.J. Hol

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Correspondence to Wim G.J. Hol.

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Ævarsson, A., Seger, K., Turley, S. et al. Crystal structure of 2-oxoisovalerate and dehydrogenase and the architecture of 2-oxo acid dehydrogenase multienzyme complexes. Nat Struct Mol Biol 6, 785–792 (1999). https://doi.org/10.1038/11563

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