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Ionization states of the catalytic residues in HIV-1 protease

Nature Structural Biology volume 3, pages 946950 (1996) | Download Citation

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Abstract

Chemical synthesis was used to prepare the HIV-1 protease specifically 13C-labelled in the catalytically essential Asp 25 in each monomer. The NMR chemical shift of the 13C-enriched homodimeric enzyme was measured in the presence of the inhibitor pepstatin, a mimic of the tetrahedral intermediate formed in enzyme catalysis. In this complex, the catalytic carboxyls do not titrate in the pH range where the enzyme is active; throughout the range pH 2.5–6.5, one Asp 25 side chain is protonated and the other deprotonated. By contrast, in the absence of inhibitor the two Asp side chains are chemically equivalent and both deprotonated at pH 6, the optimum for enzymatic activity. These direct observations of the chemical properties of the catalytic apparatus of the enzyme provide concrete information on which to base the design of improved HIV-1 protease inhibitors.

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Affiliations

  1. Department of Biochemistry, University of Queensland, Qld 4072, Australia

    • Ross Smith
  2. Center for Magnetic Resonance, University of Queensland, Qld 4072, Australia

    • Ian M. Brereton
  3. The Scripps Research Institute, La Jolla, California 92037, USA

    • Richard Y. Chai
    •  & Stephen B.H. Kent
  4. ross@biosd.uq.edu.au

    • Ross Smith
  5. stevek@chemical.ligation.com

    • Stephen B.H. Kent

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DOI

https://doi.org/10.1038/nsb1196-946

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