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A unique fold of phospholipase C-β mediates dimerization and interaction with Gαq

Nature Structural Biology volume 9, pages 32–36 (2002)Cite this article

Abstract

GTP-bound subunits of the Gq family of Gα subunits directly activate phospholipase C-β (PLC-β) isozymes to produce the second messengers inositol 1,4,5-trisphosphate and diacylglycerol. PLC-βs are GTPase activating proteins (GAPs) that also promote the formation of GDP-bound, inactive Gβ subunits. Both phospholipase activation by Gα–GTP subunits and GAP activity require a C-terminal region unique to PLC-β isozymes. The crystal structure of the C-terminal region from an avian PLC-β, determined at 2.4 Å resolution, reveals a novel fold composed almost entirely of three long helices forming a coiled-coil that dimerizes along its long axis in an antiparallel orientation. The dimer interface is extensive (∼3,200 Å2), and, based on gel exclusion chromatography, full length PLC-βs are dimeric, indicating that PLC-βs likely function as dimers. Sequence conservation, mutational data and molecular modeling show that an electrostatically positive surface of the dimer contains the major determinants for binding Gβq. Effector dimerization, as highlighted by PLC-βs, provides a viable mechanism for regulating signaling cascades linked to heterotrimeric G proteins.

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Figure 1: Overall structure of the CT domain unique to PLC-βs.
Figure 2: Dimerization of CT domains and full length PLC-β.
Figure 3: Dimeric CTtΔ is highly electrostatically polarized.
Figure 4: Biochemical and structural characterization of CT domains of PLC-βs with Gαq.

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Acknowledgements

We thank M. Pham and S. Foster for technical assistance, H. Ke and L. Pedersen (NIEHS) for help in data collection and D. Worthylake and M. Pliske for technical assistance and critical reading of the manuscript. We would also like to thank the staff at beamline 5-1 of the Stanford Synchotron Research Laboratories, a national user facility operated by Stanford University on behalf of the U.S. Department of Energy. J.S. and K.H. acknowledge support from the NIH. J.S. also acknowledges support from the Pew Charitable Trusts.

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Authors and Affiliations

  1. Department of Pharmacology, The University of North Carolina at Chapel Hill, North Carolina, 27599, USA

    Alex U. Singer, Gary L. Waldo, T. Kendall Harden & John Sondek

  2. Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, North Carolina, 27599, USA

    John Sondek

  3. Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, North Carolina, 27599, USA

    John Sondek

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  1. Alex U. Singer
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Correspondence to John Sondek.

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Singer, A., Waldo, G., Harden, T. et al. A unique fold of phospholipase C-β mediates dimerization and interaction with Gαq. Nat Struct Mol Biol 9, 32–36 (2002). https://doi.org/10.1038/nsb731

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