Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Structural basis of the α1–β subunit interaction of voltage-gated Ca2+ channels

Nature volume 429, pages 675–680 (2004)Cite this article

Abstract

High-voltage-activated Ca2+ channels are essential for diverse biological processes. They are composed of four or five subunits, including α1, α2-δ, β and γ (ref. 1). Their expression and function are critically dependent on the β-subunit, which transports α1 to the surface membrane and regulates diverse channel properties2,3,4. It is believed3,4,5,6 that the β-subunit interacts with α1 primarily through the β-interaction domain (BID), which binds directly to the α-interaction domain (AID) of α17; however, the molecular mechanism of the α1–β interaction is largely unclear. Here we report the crystal structures of the conserved core region of β3, alone and in complex with AID, and of β4 alone. The structures show that the β-subunit core contains two interacting domains: a Src homology 3 (SH3) domain and a guanylate kinase (GK) domain. The AID binds to a hydrophobic groove in the GK domain through extensive interactions, conferring extremely high affinity between α1 and β-subunits4,8. The BID is essential both for the structural integrity of and for bridging the SH3 and GK domains, but it does not participate directly in binding α1. The presence of multiple protein-interacting modules in the β-subunit opens a new dimension to its function as a multi-functional protein.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Sequence alignment and domain organization of Ca2+ channel α1 and β-subunits.
Figure 2: Crystal structure of the β3 core in complex with its α1 binding partner (AID).
Figure 3: Interactions between β3 and AID.
Figure 4: Structural comparisons of the β-subunit core.
Figure 5: Structural and functional comparisons of the β-subunit with other proteins.

Similar content being viewed by others

References

  1. Catterall, W. A. Structure and regulation of voltage-gated Ca2+ channels. Annu. Rev. Cell Dev. Biol. 16, 521–555 (2000)

    Article  CAS  Google Scholar 

  2. Birnbaumer, L. et al. Structures and functions of calcium channel beta subunits. J. Bioenerg. Biomembr. 30, 357–375 (1998)

    Article  CAS  Google Scholar 

  3. Arikkath, J. & Campbell, K. P. Auxiliary subunits: essential components of the voltage-gated calcium channel complex. Curr. Opin. Neurobiol. 13, 298–307 (2003)

    Article  CAS  Google Scholar 

  4. Dolphin, A. C. β subunits of voltage-gated calcium channels. J. Bioenerg. Biomembr. 55, 607–627 (2003)

    CAS  Google Scholar 

  5. De Waard, M., Pragnell, M. & Campbell, K. P. Ca2+ channel regulation by a conserved beta subunit domain. Neuron 13, 495–503 (1994)

    Article  CAS  Google Scholar 

  6. De Waard, M., Scott, V. E., Pragnell, M. & Campbell, K. P. Identification of critical amino acids involved in α1-β interaction in voltage-dependent Ca2+ channels. FEBS Lett. 380, 272–276 (1996)

    Article  CAS  Google Scholar 

  7. Pragnell, M. et al. Calcium channel β-subunit binds to a conserved motif in the I–II cytoplasmic linker of the α1-subunit. Nature 368, 67–70 (1994)

    Article  ADS  CAS  Google Scholar 

  8. Canti, C. et al. Evidence for two concentration-dependent processes for β-subunit effects on α1B calcium channels. Biophys. J. 81, 1439–1451 (2001)

    Article  ADS  CAS  Google Scholar 

  9. Opatowsky, Y., Chomsky-Hecht, O., Kang, M. G., Campbell, K. P. & Hirsch, J. A. The voltage-dependent calcium channel beta subunit contains two stable interacting domains. J. Biol. Chem. 278, 52323–52332 (2003)

    Article  CAS  Google Scholar 

  10. Hanlon, M. R., Berrow, N. S., Dolphin, A. C. & Wallace, B. A. Modelling of a voltage-dependent Ca2+ channel beta subunit as a basis for understanding its functional properties. FEBS Lett. 445, 366–370 (1999)

    Article  CAS  Google Scholar 

  11. Hendrickson, W. A. Determination of macromolecular structures from anomalous diffraction of synchrotron radiation. Science 254, 51–58 (1991)

    Article  ADS  CAS  Google Scholar 

  12. Tavares, G. A., Panepucci, E. H. & Brunger, A. T. Structural characterization of the intramolecular interaction between the SH3 and guanylate kinase domains of PSD-95. Mol. Cell 8, 1313–1325 (2001)

    Article  CAS  Google Scholar 

  13. Zarrinpar, A., Bhattacharyya, R. P. & Lim, W. A. The structure and function of proline recognition domains. Sci. STKE 2003, RE8 (2003)

    PubMed  Google Scholar 

  14. Larson, S. M. & Davidson, A. R. The identification of conserved interactions within the SH3 domain by alignment of sequences and structures. Protein Sci. 9, 2170–2180 (2000)

    Article  CAS  Google Scholar 

  15. Blaszcyk, J., Li, Y., Yan, H. & Ji, X. Crystal structure of unligated guanylate kinase from yeast reveals GMP-induced conformational changes. J. Mol. Biol. 307, 247–257 (2001)

    Article  Google Scholar 

  16. Sheng, M. & Pak, D. T. Ligand-gated ion channel interactions with cytoskeletal and signaling proteins. Annu. Rev. Physiol. 62, 755–778 (2000)

    Article  CAS  Google Scholar 

  17. Beguin, P. et al. Regulation of Ca2+ channel expression at the cell surface by the small G-protein kir/Gem. Nature 411, 701–706 (2001)

    Article  ADS  CAS  Google Scholar 

  18. Finlin, B. S., Crump, S. M., Satin, J. & Andres, D. A. Regulation of voltage-gated calcium channel activity by the Rem and Rad GTPases. Proc. Natl Acad. Sci. USA 100, 14469–14474 (2003)

    Article  ADS  CAS  Google Scholar 

  19. McGee, A. W. et al. Structure of the SH3-guanylate kinase module from PSD-95 suggests a mechanism for regulated assembly of MAGUK scaffolding proteins. Mol. Cell 8, 1291–1301 (2001)

    Article  CAS  Google Scholar 

  20. Garcia, E. P. et al. SAP90 binds and clusters kainate receptors causing incomplete desensitization. Neuron 21, 727–739 (1998)

    Article  CAS  Google Scholar 

  21. Maximov, A., Sudhof, T. C. & Bezprozvanny, I. Association of neuronal calcium channels with modular adaptor proteins. J. Biol. Chem. 274, 24453–24456 (1999)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  23. Terwilliger, T. C. & Berendzen, J. Automated MAD and MIR structure solution. Acta Crystallogr. D 55, 849–861 (1999)

    Article  CAS  Google Scholar 

  24. Jones, T. A., Zou, J. Y.,, Cowan, S. W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A 47, 110–119 (1991)

    Article  Google Scholar 

  25. Brunger, A. T. et al. Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr. D 54, 905–921 (1998)

    Article  CAS  Google Scholar 

  26. Jogl, G., Tao, X., Xu, Y. & Tong, L. COMO: a program for combined molecular replacement. Acta Crystallogr. D 57, 1127–1134 (2001)

    Article  CAS  Google Scholar 

  27. Wu, L., Bauer, C. S., Zhen, X. G., Xie, C. & Yang, J. Dual regulation of voltage-gated calcium channels by PtdIns(4,5)P2. Nature 419, 947–952 (2002)

    Article  ADS  CAS  Google Scholar 

  28. Carson, M. Ribbon models of macromolecules. J. Mol. Graph. 5, 103–106 (1987)

    Article  CAS  Google Scholar 

  29. Nicholls, A., Sharp, K. & Honig, B. H. Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins Struct. Funct. Genet. 11, 281–296 (1991)

    Article  CAS  Google Scholar 

  30. Kraulis, P. MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Crystallogr. 24, 946–950 (1991)

    Article  Google Scholar 

Download references

Acknowledgements

We thank Y. Mori for CaV2.1 complementary DNA; E. Perez-Reyes for all β-subunit cDNAs; T. Tanabe for α2δ cDNA; and the staff at X4A of NSLS, Brookhaven National Laboratory, for synchrotron support. This work was supported by grants to L.T. and J.Y. from the National Institutes of Health. J.Y. is a recipient of the McKnight Scholar Award, the Scholar Research Programme of the EJLB Foundation and the Established Investigator Award of the American Heart Association.

Author information

Authors and Affiliations

  1. Department of Biological Sciences, Columbia University, New York, New York, 10027, USA

    Yu-hang Chen, Ming-hui Li, Yun Zhang, Lin-ling He, Yoichi Yamada, Aileen Fitzmaurice, Yang Shen, Hailong Zhang, Liang Tong & Jian Yang

Authors
  1. Yu-hang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ming-hui Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lin-ling He
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yoichi Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Aileen Fitzmaurice
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yang Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hailong Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Liang Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Jian Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jian Yang.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure 1

Comparison of the structures of the core region of Ca2+ channel β subunits. (DOC 2325 kb)

Supplementary Figure 2

BID is not directly involved in binding AID. (DOC 784 kb)

Supplementary Figure 3

Stimulation of Ca2+ channel expression by an artificial peptide. (DOC 530 kb)

Supplementary Figure 4

Quality of structural models. (DOC 1782 kb)

Supplementary Table 1

Crystallographic Data, phasing statistics, and refinement statistics. (DOC 43 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, Yh., Li, Mh., Zhang, Y. et al. Structural basis of the α1–β subunit interaction of voltage-gated Ca2+ channels. Nature 429, 675–680 (2004). https://doi.org/10.1038/nature02641

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature02641

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing