Abstract
A multitude of heptahelical receptors use heterotrimeric G proteins to transduce signals to specific effector target molecules. The G protein transducin, Gt, couples photon-activated rhodopsin with the effector cyclic GMP phosophodiesterase (PDE) in the vertebrate phototransduction cascade. The interactions of the Gt α-subunit (αt) with the inhibitory PDE γ-subunit (PDEγ) are central to effector activation, and also enhance visual recovery in cooperation with the GTPase-activating protein regulator of G-protein signalling (RGS)-9 (refs 1,2,3). Here we describe the crystal structure at 2.0 Å of rod transducin Å·GDP·AlF-4 in complex with the effector molecule PDEγ and the GTPase-activating protein RGS9. In addition, we present the independently solved crystal structures of the RGS9 RGS domain both alone and in complex with αt/i1·GDP·AlF-4. These structures reveal insights into effector activation, synergistic GTPase acceleration, RGS9 specificity and RGS activity. Effector binding to a nucleotide-dependent site on αt sequesters PDEγ residues implicated in PDE inhibition, and potentiates recruitment of RGS9 for hydrolytic transition state stabilization and concomitant signal termination.
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Acknowledgements
We thank C. Berlot, Y. Korkhin, D. Lambright, L. Rice and J. Steitz for useful discussions; A. Brunger, P. Adams, R. Grosse-Kunstleve for help with CNS; and W. Minor and Z. Otwinowski for pre-release of HKL2000. We also thank staff of the SBC beamline 19-ID: R. Alkire, N. E.C. Duke, S. L. Ginell, K. Lazarski, S. Korolev, F. J. Rotella, R. Sanishvili, J. Lazarz, and A. Joachimiak. Use of the ANL SBC beamline at the APS was supported by the US Department of Energy, Office of Biological and Environmental Research. This work is supported in part by a NIH grant to P.B.S. M.A.K. is a HHMI Fellow of the LSRF. C.W.C. was supported by an NIH grant.
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Slep, K., Kercher, M., He, W. et al. Structural determinants for regulation of phosphodiesterase by a G protein at 2.0 Å. Nature 409, 1071–1077 (2001). https://doi.org/10.1038/35059138
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DOI: https://doi.org/10.1038/35059138
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