Abstract
The nitrophorins comprise an unusual family of proteins that use ferric (Fe(III)) heme to transport highly reactive nitric oxide (NO) from the salivary gland of a blood sucking bug to the victim, resulting in vasodilation and reduced blood coagulation. We have determined structures of nitrophorin 4 in complexes with H2O, cyanide and nitric oxide. These structures reveal a remarkable feature: the nitrophorins have a broadly open distal pocket in the absence of NO, but upon NO binding, three or more water molecules are expelled and two loops fold into the distal pocket, resulting in the packing of hydrophobic groups around the NO molecule and increased distortion of the heme. In this way, the protein apparently forms a ‘hydrophobic trap’ for the NO molecule. The structures are very accurate, ranging between 1.6 and 1.4 Å resolutions.
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References
Bredt, D.S. & Snyder, S.H. Annu. Rev. Biochem. 63, 175–195 (1994).
Law, J., Ribeiro, J.M.C. & Wells, M. Annu. Rev. Biochem. 61, 87–111 (1992).
Montfort, W.R., Weichsel, A. & Andersen, J.F. Biochim. Biophys. Acta, in the press (2000).
Kirchhoff, L.V. N. Engl. J. Med. 329, 639–644 (1993).
Sharma, V.S. & Magde, D. Methods 19, 494–505 (1999).
Ribeiro, J.M.C., Hazzard, J.M.H., Nussenzveig, R.H., Champagne, D.E. & Walker, F.A. Science 260, 539–541 (1993).
Andersen, J.F. et al. Biochemistry 36, 4423–4428 (1997).
Traylor, T.G. & Sharma, V.S. Biochemistry 31, 2847–2849 (1992).
Sharma, V.S., Isaacson, R.A., John, M.E., Waterman, M.R. & Chevion, M. Biochemsitry 22, 3897–3902 (1983).
Addison, A.W. & Stephanos, J.J. Biochemistry 25, 4104–4113 (1986).
Hoshino, M., Maeda, M., Konishi, R., Seki, H. & Ford, P.C. J. Am. Chem. Soc. 118, 5702–5707 (1996).
Andersen, J.F., Weichsel, A., Balfour, C.A., Champagne, D.E. & Montfort, W.R. Structure 6, 1315–1327 (1998).
Weichsel, A., Andersen, J.F., Champagne, D.E., Walker, F.A. & Montfort, W.R. Nature Struct. Biol. 5, 304–309 (1998).
Kaneko, Y., Yuda, M., Iio, T., Murase, T. & Chinzei, Y. Biochim. Biophys. Acta 1431, 492–499 (1999).
Sharma, V.S., Traylor, T.G., Gardiner, R. & Mizukami, H. Biochemistry 26, 3837–3843 (1987).
Ding, X.D. et al. J. Am. Chem. Soc. 121, 128–138 (1999).
Scheidt, W.R. & Ellison, M.K. Acc. Chem. Res. 32, 350–359 (1999).
Hodge, S.J. et al. Chem. Commun. 19, 2283–2284 (1996).
Edwards, S.L., Kraut, J. & Poulos, T.L. Biochemistry 27, 8074–8081 (1988).
Jewsbury, P., Yamamoto, S., Minato, T., Saito, M. & Kitagawa, T. J. Am. Chem. Soc. 116, 11586–11587 (1994).
Carducci, M.D., Pressprich, M.R. & Coppens, P. J. Am. Chem. Soc. 119, 2669–2678 (1997).
Shaw, A.W. & Vosper, A.J. J. Chem. Soc. Faraday Trans. 8, 1239–1244 (1977).
Kerwin, J.F., Jr., Lancaster, J.R., Jr. & Feldman, P.L. J. Med. Chem. 38, 4343–4362 (1995).
Walker, F.A., Emrick, D., Rivera, J.E., Hanquet, B.J. & Buttlaire, D.H. J. Am. Chem. Soc. 110, 6234–6240 (1988).
Shelnutt, J.A. et al. Chem. Soc. Rev. 27, 31–41 (1998).
Rohlfs, R.J. et al. J. Biol. Chem. 265, 3168–3176 (1990).
Messerschmidt, A. & Pflugrath, J.W. J. Appl. Crystallogr. 20, 306–315 (1987).
Kabsch, W. J. Appl. Crystallogr. 21, 916–934 (1988).
CCP4. Acta Crystallogr. D 50, 760–763 (1994).
Jones, T.A., Zou, J.Y., Cowan, S.W. & Kjelgard, M. Acta Crystallogr. A 47, 110–119 (1991).
Brunger, A.T. X-PLOR: a system for X-ray crystallography and NMR (Yale University Press, New Haven, Connecticut; 1992).
Sheldrick, G.M. & Schneider, T.R. Methods Enzymol. 277, 319–343 (1997).
Andersen, J.F. et al. Biochemistry in the press (2000).
Acknowledgements
We thank C. Balfour for protein purification. Supported in part by grants from NIH, ACS, and ADCRC to W.R.M., and from the NIH to J.F.A.
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Weichsel, A., Andersen, J., Roberts, S. et al. Nitric oxide binding to nitrophorin 4 induces complete distal pocket burial. Nat Struct Mol Biol 7, 551–554 (2000). https://doi.org/10.1038/76769
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DOI: https://doi.org/10.1038/76769
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