Abstract
The atrial natriuretic peptide (ANP) hormone is secreted by the heart in response to an increase in blood pressure. ANP exhibits several potent anti-hypertensive actions in the kidney, adrenal gland and vascular system. These actions are induced by hormone binding extracellularly to the ANP receptor1, thereby activating its intracellular guanylyl cyclase domain for the production of cyclic GMP2. Here we present the crystal structure of the glycosylated dimerized hormone-binding domain of the ANP receptor at 2.0-Å resolution. The monomer comprises two interconnected subdomains, each encompassing a central β-sheet flanked by α-helices, and exhibits the type I periplasmic binding protein fold. Dimerization is mediated by the juxtaposition of four parallel helices, arranged two by two, which brings the two protruding carboxy termini into close relative proximity. From affinity labelling and mutagenesis studies, the ANP-binding site maps to the side of the dimer crevice and extends to near the dimer interface. A conserved chloride-binding site is located in the membrane distal domain, and we found that hormone binding is chloride dependent. These studies suggest mechanisms for hormone activation and the allostery of the ANP receptor.
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Acknowledgements
This work was supported by grants from the NIH (K.S.M.) and the American Heart Association (K.S.M. and V.C.Y.) and by a NIH postdoctoral fellowship (F.v.d.A.). We thank G. Stark for his support. Diffraction data were measured at the Advanced Light Source and on the Structural Biology Center beamline at the Advanced Photon Source, supported by the US Department of Energy, and at the National Synchrotron Light Source, Brookhaven National Laboratory, supported by the U.S. Department of Energy and the National Institutes of Health. We are grateful to E. Walker and the CCF LRI Computer Core for facilities support, to S. Ginell and T. Earnest for beamline support, to L. Pearl for coordinates of the AmiC/AmiR structure, and to M. Young for interesting discussions. Purification and crystallization of the ANP receptor hormone-binding domain, ANP-binding site determination, binding studies and other functional characterizations were carried out by X.Z., M.M., X.H. and K.S.M. Data collection, structure determination and refinement, and structure analysis were carried out by F.v.d.A. and V.C.Y.
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van den Akker, F., Zhang, X., Miyagi, M. et al. Structure of the dimerized hormone-binding domain of a guanylyl- cyclase-coupled receptor. Nature 406, 101–104 (2000). https://doi.org/10.1038/35017602
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DOI: https://doi.org/10.1038/35017602
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