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A mechanism-based inactivator of glycoside hydrolases involving formation of a transient non-classical carbocation

Author

Listed:
  • Saswati Chakladar

    (Simon Fraser University)

  • Yi Wang

    (Simon Fraser University)

  • Thomas Clark

    (Simon Fraser University)

  • Lydia Cheng

    (Simon Fraser University)

  • Shirley Ko

    (Simon Fraser University)

  • David J. Vocadlo

    (Simon Fraser University
    Simon Fraser University)

  • Andrew J. Bennet

    (Simon Fraser University)

Abstract

The design of mechanism-based enzyme inactivators to generate chemical probes for biological research is an important challenge in carbohydrate chemistry. Here we describe the synthesis and biological evaluation of a novel carbocyclic mechanism-based inactivator of galactosidases (glycoside hydrolase families 27 and 36). Upon catalysis of this unnatural substrate, a transient non-classical carbocation forms within the enzyme active site. We show that the inactivation event, which proceeds via a bicyclobutonium ion intermediate, leads to a single alkylation event that occurs on the enzymatic nucleophile, an aspartic acid residue in this case. We also show that the catalytic proficiencies for enzymatic hydrolysis of substrates and inactivation by our bicyclo[4.1.0]heptyl analogue of galactose differ by only a factor of 20. This inactivator has the potential for further development as a useful biological research tool for both basic research and biotechnological applications.

Suggested Citation

  • Saswati Chakladar & Yi Wang & Thomas Clark & Lydia Cheng & Shirley Ko & David J. Vocadlo & Andrew J. Bennet, 2014. "A mechanism-based inactivator of glycoside hydrolases involving formation of a transient non-classical carbocation," Nature Communications, Nature, vol. 5(1), pages 1-8, December.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms6590
    DOI: 10.1038/ncomms6590
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