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X-ray structures along the reaction pathway of cyclodextrin glycosyltransferase elucidate catalysis in the α-amylase family

Nature Structural Biology volume 6, pages 432–436 (1999)Cite this article

Abstract

Cyclodextrin glycosyltransferase (CGTase) is an enzyme of the α-amylase family, which uses a double displacement mechanism to process α-linked glucose polymers. We have determined two X-ray structures of CGTase complexes, one with an intact substrate at 2.1 Å resolution, and the other with a covalently bound reaction intermediate at 1.8 Å resolution. These structures give evidence for substrate distortion and the covalent character of the intermediate and for the first time show, in atomic detail, how catalysis in the α-amylase family proceeds by the concerted action of all active site residues.

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Figure 1: Scheme of the CGTase reaction mechanism.
Figure 2: Stereoview of the substrate bound to CGTase.
Figure 3: Stereoview of the covalent intermediate electron density from a 2Fo - Fc omit37 map (1σ contoured), the arrow indicates the glycosyl–enzyme covalent bond.
Figure 4: Structures of the stable states along the reaction coordinate.

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Acknowledgements

Our thanks go to M. Dowd for ascertaining the true conformations of substrate and intermediate, and to T. Lindhorst for synthesizing 4-deoxymaltose. This work was supported by the EC grant entitled Structure-function relationships of glycosyltransferases.

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Authors and Affiliations

  1. BIOSON Research Institute, Groningen Biomolecular Sciences and Biotechnology Institute (GBB) and Laboratory of Biophysical Chemistry, University of Groningen, 9747 AG, Groningen, Nijenborgh, 4, The Netherlands

    Joost C. M. Uitdehaag, Kor H. Kalk & Bauke W. Dijkstra

  2. Department of Chemistry, University of British Columbia, Vancouver, V6T 1Z1, British Columbia, Canada

    Renée Mosi & Stephen G. Withers

  3. Department of Microbiology, Kerklaan 30, 9751 NN Haren, GBB Institute, University of Groningen, The Netherlands

    Bart A. van der Veen & Lubbert Dijkhuizen

Authors
  1. Joost C. M. Uitdehaag
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Uitdehaag, J., Mosi, R., Kalk, K. et al. X-ray structures along the reaction pathway of cyclodextrin glycosyltransferase elucidate catalysis in the α-amylase family. Nat Struct Mol Biol 6, 432–436 (1999). https://doi.org/10.1038/8235

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