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The aspartic proteinase from Saccharomyces cerevisiae folds its own inhibitor into a helix

Nature Structural Biology volume 7, pages 113–117 (2000)Cite this article

Abstract

Aspartic proteinase A from yeast is specifically and potently inhibited by a small protein called IA3 from Saccharomyces cerevisiae . Although this inhibitor consists of 68 residues, we show that the inhibitory activity resides within the N-terminal half of the molecule. Structures solved at 2.2 and 1.8 Å, respectively, for complexes of proteinase A with full-length IA3 and with a truncated form consisting only of residues 2–34, reveal an unprecedented mode of inhibitor–enzyme interactions. Neither form of the free inhibitor has detectable intrinsic secondary structure in solution. However, upon contact with the enzyme, residues 2–32 become ordered and adopt a near-perfect α-helical conformation. Thus, the proteinase acts as a folding template, stabilizing the helical conformation in the inhibitor, which results in the potent and specific blockage of the proteolytic activity.

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Figure 1: Schematic diagram of the structure of proteinase A.
Figure 2: Inhibitor IA3 and its interactions with proteinase A.
Figure 3: Interactions in the vicinity of the active site of proteinase A.

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Acknowledgements

This paper is dedicated to the memory of H. Holzer, formerly of the University of Freiburg im Breisgau, Germany. We thank A. Chung at the Protein Chemistry Core, University of Florida, for synthesis of peptides; A. Edison and R.A. Byrd for their valuable contributions with the NMR; V. Dhanaraj and B. Brownsey for their extensive help with modeling and mass spectroscopy analysis, respectively; J. Collins for help with the preparation of Fig. 1; and A. Arthur for editorial comments. This work was supported in part by awards from the BBSRC (ROPA) and from Actelion, Allschwil, Switzerland (J.K.), by an NIH grant (B.M.D.) and by the National Cancer Institute, DHHS, under contract with ABL (A.W.).

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Author notes

  1. Mi Li, Lowri H. Phylip and Ben M. Dunn: These authors contributed equally to this work.

Authors and Affiliations

  1. Macromolecular Crystallography Laboratory, Program in Structural Biology, National Cancer Institute-FCRDC, Frederick , 21702, Maryland, USA

    Mi Li, Alexander Wlodawer & Alla Gustchina

  2. Intramural Research Support Program, SAIC Frederick, National Cancer Institute-FCRDC, Frederick, 21702, Maryland , USA

    Mi Li

  3. School of Biosciences, Cardiff University, Cardiff, CF1 3US, UK

    Lowri H. Phylip, Wendy E. Lees & John Kay

  4. Department of Yeast Genetics, Carlsberg Laboratory, Copenhagen Valby, DK 2500, Denmark

    Jakob R. Winther

  5. Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, 32610, Florida, USA

    Ben M. Dunn

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  1. Mi Li
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Correspondence to John Kay or Alla Gustchina.

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Li, M., Phylip, L., Lees, W. et al. The aspartic proteinase from Saccharomyces cerevisiae folds its own inhibitor into a helix. Nat Struct Mol Biol 7, 113–117 (2000). https://doi.org/10.1038/72378

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