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The structure of a Michaelis serpin–protease complex

Nature Structural Biology volume 8, pages 979–983 (2001)Cite this article

Abstract

Serine protease inhibitors (serpins) regulate the activities of circulating proteases. Serpins inhibit proteases by acylating the serine hydroxyl at their active sites. Before deacylation and complete proteolysis of the serpin can occur, massive conformational changes are triggered in the serpin while maintaining the covalent linkage between the protease and serpin. Here we report the structure of a serpin–trypsin Michaelis complex, which we visualized by using the S195A trypsin mutant to prevent covalent complex formation. This encounter complex reveals a more extensive interaction surface than that present in small inhibitor–protease complexes and is a template for modeling other serpin–protease pairs. Mutations of several serpin residues at the interface reduced the inhibitory activity of the serpin. The serine residue C-terminal to the scissile peptide bond is found in a closer than usual interaction with His 57 at the active site of trypsin.

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Figure 1: Overall structure of serpin-protease complex and comparison with other serpins.
Figure 2: Stereo diagram showing the RCL between P4 and P4′ in the electron density and interactions with trypsin.
Figure 3
Figure 4: Structure-based sequence alignment of serpin RCLs.

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Acknowledgements

This research was supported by The Welch Foundation Grant (E.J.G.) and the NIH (M.R.K.). We thank S. Sprang and E. Madison for suggestions on this manuscript.

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Authors and Affiliations

  1. Department of Biochemistry, The University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Boulevard, Dallas, 75390, Texas, USA

    Sheng Ye, Amy L. Cech, Ricardo Belmares & Elizabeth J. Goldsmith

  2. Department of Pharmacology, The University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Boulevard, Dallas, 75390, Texas, USA

    Robert C. Bergstrom & David R. Corey

  3. Department of Biochemistry, Kansas State University, Manhattan, 66506, Kansas, USA

    Youren Tong & Michael R. Kanost

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Correspondence to Elizabeth J. Goldsmith.

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Ye, S., Cech, A., Belmares, R. et al. The structure of a Michaelis serpin–protease complex. Nat Struct Mol Biol 8, 979–983 (2001). https://doi.org/10.1038/nsb1101-979

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