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Carboxyl proteinase from Pseudomonas defines a novel family of subtilisin-like enzymes

Nature Structural Biology volume 8, pages 442–446 (2001)Cite this article

Abstract

The crystal structure of a pepstatin-insensitive carboxyl proteinase from Pseudomonas sp. 101 (PSCP) has been solved by single-wavelength anomalous diffraction using the absorption peak of bromide anions. Structures of the uninhibited enzyme and of complexes with an inhibitor that was either covalently or noncovalently bound were refined at 1.0–1.4 Å resolution. The structure of PSCP comprises a single compact domain with a diameter of ∼55 Å, consisting of a seven-stranded parallel β-sheet flanked on both sides by a number of helices. The fold of PSCP is a superset of the subtilisin fold, and the covalently bound inhibitor is linked to the enzyme through a serine residue. Thus, the structure of PSCP defines a novel family of serine-carboxyl proteinases (defined as MEROPS S53) with a unique catalytic triad consisting of Glu 80, Asp 84 and Ser 287.

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Figure 1: Two views of the PSCP molecule.
Figure 2: Structure-based sequence alignment of PSCP and subtilisin (PDB code 1GCI)15.
Figure 3: Chemical formulas and the electron density for the inhibitors of PSCP.
Figure 4: Residues forming the active site of PSCP in comparison with subtilisin.

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Acknowledgements

We thank A. Barrett for helpful discussions, A. Arthur for editorial comments and J. Alexandratos for help in preparation of the figures. This work was supported in part by a Grant-in-Aid for Scientific Research and a Grant-in-Aid for International Scientific Research (Joint Research) from the Ministry of Education, Science, Sports and Culture of Japan to K.O., by NIH grants to B.M.D., and in part with Federal funds from the National Cancer Institute, National Institutes of Health. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does the mention of trade names, commercial products or organizations imply endorsement by the U. S. Government.

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Authors and Affiliations

  1. Protein Structure Section, Macromolecular Crystallography Laboratory, National Cancer Institute at Frederick, Frederick, 21702, Maryland, USA

    Alexander Wlodawer, Mi Li & Alla Gustchina

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

    Mi Li

  3. Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, National Cancer Institute and NSLS, Brookhaven National Laboratory, Bldg. 725A-X9, Upton, 11973, New York, USA

    Zbigniew Dauter

  4. Department of Biosciences, School of Science and Engineering, Teikyo University, 1-1 Toyosatodai, Utsunomiya, 320-8551, Japan

    Kenichi Uchida

  5. Department of Applied Biology, Faculty of Textile Science, Kyoto Institute of Technology, Sakyo-ku, 606-8585, Kyoto, Japan

    Hiroshi Oyama & Kohei Oda

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

    Ben M. Dunn

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  1. Alexander Wlodawer
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Correspondence to Alexander Wlodawer.

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Wlodawer, A., Li, M., Dauter, Z. et al. Carboxyl proteinase from Pseudomonas defines a novel family of subtilisin-like enzymes. Nat Struct Mol Biol 8, 442–446 (2001). https://doi.org/10.1038/87610

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