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Atomic structure of the GCSF–receptor complex showing a new cytokine–receptor recognition scheme

Nature volume 401, pages 713–717 (1999)Cite this article

Abstract

Granulocyte colony-stimulating factor (GCSF) is the principal growth factor regulating the maturation, proliferation and differentiation of the precursor cells of neutrophilic granulocytes1 and is used to treat neutropenia2. GCSF is a member of the long-chain subtype of the class 1 cytokine superfamily, which includes growth hormone, erythropoietin, interleukin 6 and oncostatin M (ref. 3). Here we have determined the crystal structure of GCSF complexed to the BN–BC domains, the principal ligand-binding region of the GCSF receptor (GCSFR). The two receptor domains form a complex in a 2:2 ratio with the ligand, with a non-crystallographic pseudo-twofold axis through primarily the interdomain region and secondarily the BC domain. This structural view of a gp130-type receptor–ligand complex presents a new molecular basis for cytokine–receptor recognition.

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Figure 1: The GCSF–gs-CRH complex.
Figure 2: Secondary structures and sequences of GCSF and gs-CRH.
Figure 3: Structural comparison of the GCSF–gs-CRH complex and ligand-free gp130.

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References

  1. Metcalf,D. The molecular control of cell division, differentiation commitment and maturation in haemopoietic cells. Nature 339, 27–30 (1989).

    Article  ADS  CAS  Google Scholar 

  2. Welte,K. et al. Filgrastim (r-metHuG-CSF): the first 10 years. Blood 88, 1907–1929 (1996).

    CAS  PubMed  Google Scholar 

  3. Wells,J. A. & de Vos,A. M. Hematopoietic receptor complexes. Annu. Rev. Biochem. 65, 609–634 (1996).

    Article  CAS  Google Scholar 

  4. Fukunaga,R., Ishizaka-Ikeda,E., Pan, C.-X., Seto,Y. & Nagata,S. Functional domains of the granulocyte colony-stimulating factor receptor. EMBO J. 10, 2855–2865 (1991).

    Article  CAS  Google Scholar 

  5. Hiraoka,O., Anaguchi,H. & Ota,Y. Formation of 1:1 complex of the cytokine receptor homologous region of granulocyte colony-stimulating factor receptor with ligand. Biosci. Biotechnol. Biochem. 59, 2351–2354 (1995).

    Article  CAS  Google Scholar 

  6. Hiraoka,O., Anaguchi,H., Asakura,A. & Ota,Y. Requirement for the immunoglobulin-like domain of granulocyte colony-stimulating factor receptor in formation of a 2:1 receptor–ligand complex. J. Biol. Chem. 270, 25928–25934 (1995).

    Article  CAS  Google Scholar 

  7. Horan,T. P. et al. Coexpression of G-CSF with an unglycosylated G-CSF receptor mutant results in secretion of a stable complex. Protein Express. Purific. 14, 45–53 (1998).

    Article  CAS  Google Scholar 

  8. Hiraoka,O., Anaguchi,H. & Ota,Y. Evidence for the ligand-induced conversion from a dimer to a tetramer of the granulocyte colony-stimulating factor receptor. FEBS Lett. 356, 255–260 (1994).

    Article  CAS  Google Scholar 

  9. Horan,T. P. et al. Dimerization of the extracellular domain of granulocyte-colony stimulating factor receptor by ligand binding: a monovalent ligand induces 2:2 complexes. Biochemistry 35, 4886–4896 (1996).

    Article  CAS  Google Scholar 

  10. de Vos,A. M., Ultsch,M. & Kossiakoff,A. A. Human growth hormone and extracellular domain of its receptor: crystal structure of the complex. Science 255, 306–312 (1992).

    Article  ADS  CAS  Google Scholar 

  11. Syed,R. S. et al. Efficiency of signaling through cytokine receptors depends critically on receptor orientation. Nature 395, 511–516 (1998).

    Article  ADS  CAS  Google Scholar 

  12. Hill,C. P., Osslund,T. D. & Eisenberg,D. The structure of granulocyte-colony-stimulating factor and its relationship to other growth factors. Proc. Natl Acad. Sci. USA 90, 5167–5171 (1993).

    Article  ADS  CAS  Google Scholar 

  13. Yamasaki,K., Naito,S., Anaguchi,H., Ohkubo,T. & Ota,Y. Solution structure of an extracellular domain containing the WSXWS motif of the granulocyte colony-stimulating factor receptor and its interaction with ligand. Nature Struct. Biol. 4, 498–504 (1997).

    Article  CAS  Google Scholar 

  14. Bravo,J., Staunton,D., Heath,J. K. & Jones,E. Y. Crystal structure of a cytokine-binding region of gp130. EMBO J. 17, 1665–1674 (1998).

    Article  CAS  Google Scholar 

  15. Hage,T., Sebald,W. & Reinemer,P. Crystal structure of the Interleukin-4/Receptor α chain complex reveals a mosaic binding interface. Cell 97, 271–281 (1999).

    Article  CAS  Google Scholar 

  16. Reidhaar-Olson,J. F., De Souza-Hart,J. A. & Selick,H. E. Identification of residues critical to the activity of human granulocyte colony-stimulating factor. Biochemistry 35, 9034–9041 (1996).

    Article  CAS  Google Scholar 

  17. Layton,J. E., Iaria,J., Smith,D. K. & Treutlein,H. R. Identification of a ligand-binding site on the granulocyte colony-stimulating factor receptor by molecular modeling and mutagenesis. J. Biol. Chem. 272, 29735–29741 (1997).

    Article  CAS  Google Scholar 

  18. Clackson,T. & Wells,J. A. A hot spot of binding energy in a hormone–receptor interface. Science 267, 383–386 (1995).

    Article  ADS  CAS  Google Scholar 

  19. Wells,J. A. Binding in the growth hormone receptor complex. Proc. Natl Acad. Sci. USA 93, 1–6 (1996).

    Article  ADS  CAS  Google Scholar 

  20. Hendsch,Z. S. & Tidor,B. Do salt bridges stabilize proteins? A continuum electrostatic analysis. Protein Sci. 3, 211–226 (1994).

    Article  CAS  Google Scholar 

  21. Braden,B. C. & Poljak,R. J. Structural features of the reactions between antibodies and protein antigens. FASEB J. 9, 9–16 (1995).

    Article  CAS  Google Scholar 

  22. Covell,D. G. & Wallqvist,A. Analysis of protein–protein interactions and the effects of amino acid mutations on their energetics: the importance of water molecules in the binding epitope. J. Mol. Biol. 269, 281–297 (1997).

    Article  CAS  Google Scholar 

  23. Kuga,T. et al. Japanese Patent Publication (Kokoku) 8317 Feb. 2 (1994).

  24. Yamasaki,M., Konishi,N., Yamaguchi,K., Itoh,S. & Yokoo,Y. Purification and characterization of recombinant human granulocyte colony-stimulating factor (rhG-CSF) derivatives: KW-2228 and other derivatives. Biosci. Biotechnol. Biochem. 62, 1528–1534 (1998).

    Article  CAS  Google Scholar 

  25. Livnah,O. et al. An antagonist peptide–EPO receptor complex suggests that receptor dimerization is not sufficient for activation. Nature Struct. Biol. 5, 993–1004 (1998).

    Article  CAS  Google Scholar 

  26. Livnah,O. et al. Crystallographic evidence for preformed dimers of erythropoietin receptor before ligand activation. Science 283, 987–990 (1999).

    Article  ADS  CAS  Google Scholar 

  27. Remy,I., Wilson,I. A. & Michnick,S. W. Erythropoietin receptor activation by a ligand-induced conformation change. Science 283, 990–993 (1999).

    Article  ADS  CAS  Google Scholar 

  28. Otwinowski,Z. & Minor,W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).

    Article  CAS  Google Scholar 

  29. Collaborative Computational Project, Number 4. The CCP4 Suite: Programs for crystallography. Acta Crystallogr. D 50, 760–763 (1994).

    Article  Google Scholar 

  30. Brünger,A. T. X-PLOR, Version 3.1, A System for X-ray Crystallography and NMR (Yale Univ. Press, New Haven, 1992).

    Google Scholar 

Download references

Acknowledgements

We thank N. Kamiya and M. Kawamoto for their help in the use of the data collection facilities at SPring-8, Hyogo, Japan, and N. Sakabe and N. Watanabe for use of the facilities at Photon Factory, Tsukuba, Japan. We thank S. Nagata for providing the GCSFR cDNA, E. Yanagihara and N. Okitsu for assistance in sample preparation, and Y. Kato, D. Kohda, H. Toh, K. Yamasaki and S. Tsutakawa for useful discussions. This study was partly supported by a grant from TARA (Tsukuba Advanced Research Alliance), University of Tsukuba, Japan.

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Author notes

  1. Masaharu Aritomi

    Present address: Asahi Chemical Industry Co. Ltd, 2-1 Samejima, Fuji-city, Shizuoka, 416-8501, Japan

  2. Yoshimi Ota

    Present address: Toray Industrial, Inc., 1111 Tebiro, Kamakura-city, Kanagawa, 248-0036, Japan

Authors and Affiliations

  1. Biomolecular Engineering Research Institute, 6-2-3 Furuedai, Suita-city, 565-0874, Osaka, Japan

    Masaharu Aritomi, Naoki Kunishima, Tomoyuki Okamoto, Yoshimi Ota & Kosuke Morikawa

  2. Central Laboratories for Key Technology, Kirin Brewery Co. Ltd, 13-5 Fukuura, Kanazawa-ku, Yokohama-city, 236-0004, Kanagawa, Japan

    Tomoyuki Okamoto & Ryota Kuroki

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Correspondence to Kosuke Morikawa.

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Aritomi, M., Kunishima, N., Okamoto, T. et al. Atomic structure of the GCSF–receptor complex showing a new cytokine–receptor recognition scheme. Nature 401, 713–717 (1999). https://doi.org/10.1038/44394

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