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Functional and structural characterization of a heparanase

Nature Chemical Biology volume 11, pages 955–957 (2015)Cite this article

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Abstract

We report the structural and functional characterization of a novel heparanase (BpHep) from the invasive pathogenic bacterium Burkholderia pseudomallei (Bp), showing ∼24% sequence identity with human heparanase (hHep). Site-directed mutagenesis studies confirmed the active site resi-dues essential for activity, and we found that BpHep has specificity for heparan sulfate. Finally, we describe the first heparanase X-ray crystal structure, which provides new insight into both substrate recognition and inhibitor design.

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Figure 1: Characterization of B. pseudomallei heparanase (BpHep).
Figure 2: Analysis of B. pseudomallei heparanase (BpHepwt) and human heparanase cleavage of 1.
Figure 3: Crystallographic structure of B. pseudomallei heparanase (BpHepwt, PDB ID: 5BWI).

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Protein Data Bank

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Acknowledgements

We gratefully acknowledge financial support from the Rosebud Foundation and Australian Synchrotron scientists for their support during X-ray diffraction data collection at this facility's beamlines, MX1 and MX2. We thank B. Matthews, Smart Water Research Centre (Griffith University), for excellent mass spectrometry technical support.

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Author notes

  1. Lisa Bohlmann, Gregory D Tredwell and Xing Yu: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Glycomics, Griffith University, Gold Coast Campus, Southport, Queensland, Australia

    Lisa Bohlmann, Gregory D Tredwell, Xing Yu, Chih-Wei Chang, Thomas Haselhorst, Moritz Winger, Jeffrey C Dyason, Robin J Thomson, Joe Tiralongo, Ifor R Beacham, Helen Blanchard & Mark von Itzstein

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  1. Lisa Bohlmann
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Contributions

L.B., G.D.T., J.T., R.J.T., I.R.B. and M.v.I. contributed to molecular biology, protein chemistry and data analysis; L.B., C.-W.C., T.H., I.R.B. and M.v.I. contributed to NMR experiments and data analysis; C.-W.C. synthesized HS fragments; L.B., X.Y., H.B. and M.v.I. contributed to protein crystallization and structure determination; L.B., J.C.D., M.W., X.Y., H.B. and M.v.I. contributed to molecular modeling; all authors contributed to manuscript preparation; M.v.I. conceived the study.

Corresponding authors

Correspondence to Ifor R Beacham, Helen Blanchard or Mark von Itzstein.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Table 1 and Supplementary Figures 1–13. (PDF 13224 kb)

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Bohlmann, L., Tredwell, G., Yu, X. et al. Functional and structural characterization of a heparanase. Nat Chem Biol 11, 955–957 (2015). https://doi.org/10.1038/nchembio.1956

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