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Structure of outer membrane protein A transmembrane domain by NMR spectroscopy

Nature Structural Biology volume 8, pages 334–338 (2001)Cite this article

Abstract

We have determined the three-dimensional fold of the 19 kDa (177 residues) transmembrane domain of the outer membrane protein A of Escherichia coli in dodecylphosphocholine (DPC) micelles in solution using heteronuclear NMR. The structure consists of an eight-stranded β-barrel connected by tight turns on the periplasmic side and larger mobile loops on the extracellular side. The solution structure of the barrel in DPC micelles is similar to that in n-octyltetraoxyethylene (C8E4) micelles determined by X-ray diffraction. Moreover, data from NMR dynamic experiments reveal a gradient of conformational flexibility in the structure that may contribute to the membrane channel function of this protein.

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Figure 1: TROSY-based 1H-15N HSQC spectrum of 1 mM OmpA(0–176) in 600 mM deuterated DPC micelles in 10 mM potassium phosphate buffer, pH 6.3 (containing 50 mM NaCl, and 0.1 % NaN3).
Figure 2: Sequential assignment of OmpA(0–176).
Figure 3: Solution structure of OmpA TM domain in detergent micelles.
Figure 4: Dynamics of OmpA TM domain in detergent micelles.

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References

  1. Bax, A. Curr. Op. Struct. Biol. 4, 738–744 (1994).

    Article  CAS  Google Scholar 

  2. Gardner, K. H. & Kay, L.E. Annu. Rev. Biomol. Struct . 27, 357–406 (1998).

    Article  CAS  Google Scholar 

  3. MacKenzie, K.R., Prestegard J.H. & Engelman, D.M. Science 276, 131–3 (1997).

    Article  Google Scholar 

  4. Rastogi, V.K. & Girvin, M.E. Nature 402, 263–8 (1999).

    Article  CAS  Google Scholar 

  5. Ketchem, R.R., Hu, W. & Cross, T.A. Science 261, 1457–1460 (1993).

    Article  CAS  Google Scholar 

  6. Opella, S.J. et al. Nature Struct. Biol. 6, 374–379 (1999).

    Article  CAS  Google Scholar 

  7. Pervushin, K., Riek, R., Wider, G. & Wüthrich, K. Proc. Natl. Acad. Sci. USA 94, 12366–12371 (1997).

    Article  CAS  Google Scholar 

  8. Kay, L.E. & Gardner, K.H. Curr. Opin. Struct. Biol. 7, 722–731 (1997).

    Article  CAS  Google Scholar 

  9. Goto, N.K. & Kay, L.E. Curr. Opin. Struct. Biol. 10, 585–592 (2000).

    Article  CAS  Google Scholar 

  10. Tjandra, N. & Bax, A. Science 278, 1111–1114 (1997).

    Article  CAS  Google Scholar 

  11. Battiste, J.L. & Wagner, G. Biochemistry 39, 5355–5365 (2000).

    Article  CAS  Google Scholar 

  12. Grisshammer, R., Averbeck, P. & Sohal, A.K. Biochem. Soc. Trans. 27, 899–903 (1999).

    Article  CAS  Google Scholar 

  13. Curran, A.R., Templer, R.H. & Booth, P.L. Biochemistry 38, 9328–9336 (1999).

    Article  CAS  Google Scholar 

  14. Sanders, C.R. & Nagy, J.K. Curr. Opin. Struct. Biol. 10, 438–442 (2000).

    Article  CAS  Google Scholar 

  15. Kleinschmidt, J.H., den Blaauwen, T., Driessen, A.J.M. & Tamm, L.K. Biochemistry 38, 5006–5016 (1999).

    Article  CAS  Google Scholar 

  16. Kleinschmidt, J.H., Wiener, M. & Tamm, L.K. Protein Sci. 8, 2065–2071 (1999).

    Article  CAS  Google Scholar 

  17. Arora, A., Rinehart, D., Szabo, G. & Tamm, L.K. J. Biol. Chem. 275, 1594–1600 (2000).

    Article  CAS  Google Scholar 

  18. Saint, N., El Hamel, C., De, E. & Molle, G. FEMS Microbiol. Lett. 190, 261–265 (2000).

    Article  CAS  Google Scholar 

  19. Belaaouaj, A., Kim, K.S. & Shapiro, S.D. Science 289, 1185–1188 (2000).

    Article  CAS  Google Scholar 

  20. Soulas, C. et al. J. Immunol. 165, 2335–2340 (2000).

    Article  CAS  Google Scholar 

  21. Pautsch, A. & Schulz, G.E. Nature Struct. Biol. 5, 1013–1017 (1998).

    Article  CAS  Google Scholar 

  22. Pautsch, A. & Schulz, G.E. J. Mol. Biol. 298, 273–282 (2000).

    Article  CAS  Google Scholar 

  23. Schweizer, M., Hindennach, I., Garten, W. & Henning, U. Eur. J. Biochem. 82, 211–217 (1978).

    Article  CAS  Google Scholar 

  24. Dornmair, K., Kiefer, H. & Jähnig, F. J. Biol. Chem. 265, 18907–18911 (1990).

    CAS  PubMed  Google Scholar 

  25. Yang, D. & Kay, L.E. J. Biomol. NMR 13, 3–9 (1999).

    Article  CAS  Google Scholar 

  26. Yang, D. & Kay, L.E. J. Am. Chem. Soc. 121, 2571–2575 (1999).

    Article  CAS  Google Scholar 

  27. Powers, R. et al. J. Mol. Biol. 221, 1081–1090 (1991).

    Article  CAS  Google Scholar 

  28. Zhu, G., Kong, X. M., Sze, K.H. J. Biomol. NMR 13, 77–81 (1999).

    Article  CAS  Google Scholar 

  29. Ikura, M., Bax, A., Clore, G.M. & Gronenborn, A.M. J. Am. Chem. Soc. 112, 9020–9022 (1990).

    Article  CAS  Google Scholar 

  30. Liu, W.M. J. Mol. Biol. 275, 541–545 (1998).

    Article  CAS  Google Scholar 

  31. Caffrey, M. et al. J. Magn. Reson. 135, 368–372 (1998).

    Article  CAS  Google Scholar 

  32. Vogel, H., Nilsson, L., Rigler, R., Voges, K.P. & Jung, G. Proc. Natl. Acad. Sci. USA 85, 5067–5071 (1988).

    Article  CAS  Google Scholar 

  33. Brown, M.F., Seelig, J. & Häberlen, U. J. Chem. Phys. 70, 5045–5053 (1979).

    Article  CAS  Google Scholar 

  34. Waugh, D. J. Biomol. NMR 8, 184–192 (1996).

    Article  CAS  Google Scholar 

  35. Muchmore, D.C., McIntosh, L.P., Russell, C.B., Anderson, D.E. & Dahlquist, F.W. Methods Enzymol. 177, 44–73 (1989).

    Article  CAS  Google Scholar 

  36. Cornilescu, G., Delaglio, F. & Bax, A. J. Biomol. NMR, 13, 289–302 (1999).

    Article  CAS  Google Scholar 

  37. Gardner K.H., Rosen, M.K. & Kay, L.E. Biochemistry 36, 1389–1401 (1997).

    Article  CAS  Google Scholar 

  38. Güntert, P., Mumenthaler, C. & Wüthrich, K. J. Mol. Biol . 273, 283–298 (1997).

    Article  Google Scholar 

  39. Luginbühl, P., Güntert, P., Billeter, M. & Wüthrich, K. J. Biomol. NMR 8, 136–146 (1996).

    Article  Google Scholar 

  40. Koradi, R., Billeter, M. & Wüthrich, K. J. Mol. Graphics 14, 51–55 (1996).

    Article  CAS  Google Scholar 

  41. Farrow, N.A. et al. Biochemistry 33, 5984–6003 (2000).

    Article  Google Scholar 

  42. Laskowski, R.A., Rullmann, J.A.C., MacArthur, M.W., Kaptein, R. & Thornton, J.M. J. Biomol. NMR 8, 477–486 (1996).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by a grant from the NIH. The experiments at 750 MHz were carried out at the National Magnetic Resonance Facility at Madison, which is supported by the NIH and NSF.

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Authors and Affiliations

  1. Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, 22908-0736, Virginia, USA

    Ashish Arora, John H. Bushweller & Lukas K. Tamm

  2. NMRFAM, University of Wisconsin, Madison, 53706, Wisconsin, USA

    Frits Abildgaard

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  1. Ashish Arora
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  2. Frits Abildgaard
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  4. Lukas K. Tamm
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Correspondence to Lukas K. Tamm.

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Arora, A., Abildgaard, F., Bushweller, J. et al. Structure of outer membrane protein A transmembrane domain by NMR spectroscopy. Nat Struct Mol Biol 8, 334–338 (2001). https://doi.org/10.1038/86214

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