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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Parish, C.R., Freeman, C. & Hulett, M. Biochim. Biophys. Acta 1471, M99–M108 (2001).
Ilan, N., Elkin, M. & Vlodavsky, I. Int. J. Biochem. Cell Biol. 38, 2018–2039 (2006).
Meirovitz, A. et al. FEBS J. 280, 2307–2319 (2013).
Arvatz, G., Shafat, I., Levy-Adam, F., Ilan, N. & Vlodavsky, I. Cancer Metastasis Rev. 30, 253–268 (2011).
Höök, M., Wasteson, A. & Oldberg, A. Biochem. Biophys. Res. Commun. 67, 1422–1428 (1975).
Vlodavsky, I. et al. Semin. Cancer Biol. 12, 121–129 (2002).
Sanderson, R.D. & Iozzo, R.V. Matrix Biol. 31, 283–284 (2012).
Ziolkowski, A.F., Popp, S.K., Freeman, C., Parish, C.R. & Simeonovic, C.J. J. Clin. Invest. 122, 132–141 (2012).
Hulett, M.D. et al. Biochemistry 39, 15659–15667 (2000).
Zhou, Z., Bates, M. & Madura, J.D. Proteins 65, 580–592 (2006).
Sapay, N., Cabannes, É., Petitou, M. & Imberty, A. Biopolymers 97, 21–34 (2012).
Gandhi, N.S., Freeman, C., Parish, C.R. & Mancera, R.L. Glycobiology 22, 35–55 (2012).
Vinader, V., Haji-Abdullahi, M.H., Patterson, L.H. & Afarinkia, K. PLoS ONE 8, e82111 (2013).
Linhardt, R.J., Galliher, P.M. & Cooney, C.L. Appl. Biochem. Biotechnol. 12, 135–176 (1986).
Lohse, D.L. & Linhardt, R.J. J. Biol. Chem. 267, 24347–24355 (1992).
St John, J.A. et al. MBio 5, e00025 (2014).
Vreys, V. & David, G. J. Cell. Mol. Med. 11, 427–452 (2007).
Huang, K.-S. et al. Anal. Biochem. 333, 389–398 (2004).
Bohlmann, L., Chang, C.-W., Beacham, I.R. & von Itzstein, M. ChemBioChem 16, 1205–1211 (2015).
Hu, Y.-P. et al. J. Am. Chem. Soc. 134, 20722–20727 (2012).
Gong, F. et al. J. Biol. Chem. 278, 35152–35158 (2003).
Shimoi, K. & Nakayama, T. Methods Enzymol. 400, 263–272 (2005).
Pearson, A.G., Kiefel, M.J., Ferro, V. & von Itzstein, M. Org. Biomol. Chem. 9, 4614–4625 (2011).
Michikawa, M. et al. J. Biol. Chem. 287, 14069–14077 (2012).
Fairbanks, M.B. et al. J. Biol. Chem. 274, 29587–29590 (1999).
Holden, M.T. et al. Proc. Natl. Acad. Sci. USA 101, 14240–14245 (2004).
Battye, T.G.G., Kontogiannis, L., Johnson, O., Powell, H.R. & Leslie, A.G.W. Acta Crystallogr. D Biol. Crystallogr. 67, 271–281 (2011).
Winn, M.D. et al. Acta Crystallogr. D Biol. Crystallogr. 67, 235–242 (2011).
Evans, P. Acta Crystallogr. D Biol. Crystallogr. 62, 72–82 (2006).
McCoy, A.J. et al. J. Appl. Crystallogr. 40, 658–674 (2007).
Murshudov, G.N., Vagin, A.A. & Dodson, E.J. Acta Crystallogr. D Biol. Crystallogr. 53, 240–255 (1997).
Emsley, P. & Cowtan, K. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).
Vaguine, A.A., Richelle, J. & Wodak, S.J. Acta Crystallogr. D Biol. Crystallogr. 55, 191–205 (1999).
Laskowski, R.A., MacArthur, M.W., Moss, D.S. & Thornton, J.M. Appl. Crystallogr. 26, 283–291 (1993).
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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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.
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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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DOI: https://doi.org/10.1038/nchembio.1956
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