Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Crystal structure of brefeldin A esterase, a bacterial homolog of the mammalian hormone-sensitive lipase

Nature Structural Biology volume 6, pages 340–345 (1999)Cite this article

Abstract

Brefeldin A esterase (BFAE), a detoxifying enzyme isolated from Bacillus subtilis, hydrolyzes and inactivates BFA, a potent fungal inhibitor of intracellular vesicle-dependent secretory transport and poliovirus RNA replication. We have solved the crystal structure of BFAE and we discovered that the previously reported amino acid sequence was in serious error due to frame shifts in the cDNA sequence. The correct sequence, inferred from the experimentally phased electron density map, revealed that BFAE is a homolog of the mammalian hormone sensitive lipase (HSL). It is a canonical α/β hydrolase with two insertions forming the substrate binding pocket. The enzyme contains a lipase-like catalytic triad, Ser 202, Asp 308 and His 338, consistent with mutational studies that implicate the homologous Ser 424, Asp 693 and His 723 in the catalytic triad in human HSL.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: a, Experimentally phased electron density map in the region of the putative catalytic site.
Figure 2: a, Overview of the tertiary architecture of BFAE:
Figure 3: a, The vicinity of the catalytic site in BFAE.
Figure 4: a, Structure-aided sequence alignment of amino acid sequences of several representative members of the HSL family.

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

Protein Data Bank

References

  1. Ollis, D.L. et al. Protein Eng. 5, 597– 611 (1992).

    Article  Google Scholar 

  2. Sussman, J.L. et al. Science. 253, 872– 879 (1991).

    Article  CAS  Google Scholar 

  3. Derewenda, Z.S. Adv. Prot. Chem. 45, 1–52 (1994).

    CAS  Google Scholar 

  4. Hecht, H.J., Sobek, H., Haag, T., Pfeifer, O. & van Pee, K.H. Nature Struct. Biol. 1, 532 –537 (1994).

    Article  CAS  Google Scholar 

  5. Wagner, U.G., Hasslacher, M., Griengl, H., Schwab, H. & Kratky, C. Structure. 4, 811–22 (1996).

    Article  CAS  Google Scholar 

  6. Langin, D., Holm, C. & Lafontan, M. Proc. Nutr. Soc. 55, 93– 109 (1996).

    Article  CAS  Google Scholar 

  7. Anthonsen, M.W., Ronnstrand, L., Wernstedt, C., Degerman, E. & Holm, C. J. Biol. Chem. 273, 215–221 (1998).

    Article  CAS  Google Scholar 

  8. Holm, C., Davis, R.C., Osterlund, T., Schotz, M.C. & Fredrikson, G. FEBS Lett. 344, 234–238 (1994).

    Article  CAS  Google Scholar 

  9. Contreras, J.A. et al. J. Biol. Chem. 271, 31426– 31430 (1996).

    Article  CAS  Google Scholar 

  10. Osterlund, T., Contreras, J.A. & Holm, C. FEBS Lett. 403, 259– 262 (1997).

    Article  CAS  Google Scholar 

  11. Sugai, M., Chen, C.H. & Wu, H.C. Proc. Natl. Acad. Sci. USA 89, 8903–8907 (1992).

    Article  CAS  Google Scholar 

  12. Cuconati, A., Molla, A. & Wimmer, E. J Virol. 72, 6456– 6464 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Kneusel, R.E., Schiltz, E. & Matern, U. J. Biol. Chem. 269, 3449– 3456 (1994).

    CAS  PubMed  Google Scholar 

  14. Hemila, H., Koivula, T.T. & Palva, I. Biochim. Biophys. Acta. 1210, 249–253 (1994).

    Article  CAS  Google Scholar 

  15. Rathbone, D.A., Holt, P.J., Lowe, C.R. & Bruce, N.C. Appl. Environ. Microbiol. 63, 2062–2066 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Choo, D.W., Kurihara, T., Suzuki, T., Soda, K. & Esaki, N. Appl Environ Microbiol. 64, 486–491 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Philipp, W.J. et al. Proc. Natl. Acad. Sci. USA 93, 3132 –3137.

  18. Klenk, H.P. et al. (1997) Nature 390, 364– 370 (1996).

    Article  Google Scholar 

  19. Baudouin, E. et al. Eur. J. Biochem. 248, 700– 706 (1997).

    Article  CAS  Google Scholar 

  20. Laskowski, R.A., McArthur, M.W., Moss, D.S. & Thornton, J.M. J. Appl. Crystallogr. 26, 282–291 (1993).

    Article  Google Scholar 

  21. Derewenda, Z.S. & Wei, Y. J. Am. Chem. Soc. 117, 2104–2105.

  22. Weber, H.P., Hauser, D. & Sigg, H.P. (1971) Helvetica Chimica Acta 54, 2763–2766 (1995).

    Article  Google Scholar 

  23. Osterlund, T. et al. Biochem. J. 319, 411– 420 (1996).

    Article  CAS  Google Scholar 

  24. Wei, Y., Swenson, L., Kneusel, R.E., Matern, U. & Derewenda, Z.S. Acta Crystallogr. D 52, 1194–1195 (1996).

    Article  CAS  Google Scholar 

  25. Collaborative Computing Project Number 4. Acta Crystallogr. D 50, 760–763 (1994).

  26. Otwinowski, Z. & Minor, W. Methods. Enzymol. 276, 307–326 ( 1997).

    Article  CAS  Google Scholar 

  27. Horton, R.M. & Pease, L.R. McPherson, M.J. In Directed mutagenesis: a practical approach (ed. McPherson, M.J.) 217–247 (IRL Press, Oxford; 1991).

    Google Scholar 

  28. Brunger, A. X-PLOR, version 3.1 A system for X-ray crystallography and NMR. (Yale Univ. Press, New Haven, Connecticut; 1992).

  29. Murshudov, G.N., Vagin, A.A. & Dodson, E.J. Refinement of macromolecular structures by the maximum likelihood method. Acta Crystallogr. D53, 240–255 (1997).

    CAS  Google Scholar 

Download references

Acknowledgements

We thank the Swedish Medical Research Council and the Albert PÂhlssons Foundation for financial support

Author information

Author notes

  1. Yunyi Wei

    Present address: Vertex Pharmaceuticals Incorporated, 130 Waverly Street, Cambridge, Massachusetts, 02139, USA

Authors and Affiliations

  1. Department of Molecular Physiology and Biological Physics, University of Virginia, P.O. BOX 1011, Charlottesville , 22906-0011, Virginia, USA

    Yunyi Wei, Peter Sheffield, Urszula Derewenda & Zygmunt S. Derewenda

  2. Department of Cell and Molecular Biology, Section for Molecular Signaling, Lund University, P.O. Box 94, Lund, S-221 00, Sweden

    Juan Antonio Contreras, Torben Osterlund & Cecilia Holm

  3. Biologisches Institut II, Lehrstuhl fuer Biochemie der Planzen, Schaenzlestrasse 1, Freiburg, D-79104, Germany

    R.E. Kneusel & Ulrich Matern

  4. Qiagen GmbH, Max-Volmer-Strasse 4, Hilden, D-40724, Germany

    R.E. Kneusel

  5. Department of Pharmaceutical Biology, Marburg University, Postfach, Marburg, D-35032, Germany

    Ulrich Matern

Authors
  1. Yunyi Wei
    View author publications

    Search author on:PubMed Google Scholar

  2. Juan Antonio Contreras
    View author publications

    Search author on:PubMed Google Scholar

  3. Peter Sheffield
    View author publications

    Search author on:PubMed Google Scholar

  4. Torben Osterlund
    View author publications

    Search author on:PubMed Google Scholar

  5. Urszula Derewenda
    View author publications

    Search author on:PubMed Google Scholar

  6. R.E. Kneusel
    View author publications

    Search author on:PubMed Google Scholar

  7. Ulrich Matern
    View author publications

    Search author on:PubMed Google Scholar

  8. Cecilia Holm
    View author publications

    Search author on:PubMed Google Scholar

  9. Zygmunt S. Derewenda
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Zygmunt S. Derewenda.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wei, Y., Contreras, J., Sheffield, P. et al. Crystal structure of brefeldin A esterase, a bacterial homolog of the mammalian hormone-sensitive lipase. Nat Struct Mol Biol 6, 340–345 (1999). https://doi.org/10.1038/7576

Download citation

  • Received:

  • Accepted:

  • Issue date:

  • DOI: https://doi.org/10.1038/7576

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing